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Sample records for gemcitabine-loaded peg-pdlla nanovesicles

  1. Preparation,physicochemical characterization and cytotoxicity in vitro of gemcitabine-loaded PEG-PDLLA nanovesicles

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    AIM:To investigate the preparation,physicochemical characterization and cytotoxicity in vitro of Gemcitabine-loaded poly(ethylene glycol)-block-poly(D,L-lactide) (PEG-PDLLA) nanovesicles. METHODS:The nanovesicle carriers were prepared from the amphiphilic block copolymer of PEG-PDLLA by a double emulsion technique,and gemcitabine was used as the model drug. The morphology of the nanovesicles was determined by scanning and transmission electron microscopy,and the drug content,drug entrapment and drug-release...

  2. Synthesis, characterization and evaluation of tinidazole-loaded mPEG-PDLLA (10/90) in situ gel forming system for periodontitis treatment.

    Science.gov (United States)

    Tian, Yu; Shen, Yan; Jv, Minli

    2016-10-01

    Traditional in situ gel forming systems are potential applications for parenteral administration but always accompanied with burst release. To overcome this limitation, the tinidazole (TNZ)-loaded in situ gel forming system using a diblock copolymer, monomethoxy poly(ethylene glycol)-block-poly(d,l-lactide) (mPEG-PDLLA), was designed. The formulation of the mPEG-PDLLA-based TNZ in situ gel forming system contained 5% (w/w) TNZ, 0.4% glycerol, 5 ml N-methyl pyrrolidone (NMP) and 35% (w/w) mPEG-PDLLA. The in situ gel forming system showed sustained TNZ release over 192 h with low burst effect (around 7% in the first 8 h) in the in vitro release study. Additionally, in vivo studies were performed on rabbits with ligature-induced periodontitis, and the concentration of TNZ in the gingival crevicular fluid (GCF) as well as the pharmacokinetic parameters was calculated and the pharmacological effect of TNZ-loaded in situ gel forming (mPEG-PDLLA)-based system was found effective. Finally, histological studies revealed that the gel was a safe formulation with low irritation. The desirable drug release kinetics combined with the excellent in vivo characteristics highlight the potential of the gel in the treatment of periodontitis. Therefore, these results confirmed that the TNZ-loaded in situ gel forming mPEG-PDLLA-based system could reduce burst release of TNZ and act as a sustained-release and injectable drug depot for periodontitis treatment.

  3. Solidified mPEG-PDLLA micelles as a novel oral delivery system of indomethacin%可固化mPEG-PDLLA胶束作为载体的新型吲哚美辛口服给药系统的体外研究

    Institute of Scientific and Technical Information of China (English)

    Ammar Ouahab; 沈雁; 平其能; 涂家生

    2011-01-01

    本研究旨在制备吲哚美辛固化聚合物胶束(IND-SPM),并研究其体外释放特性.IND-SPM以交联聚维酮XL-10作为载体,并以聚乙二醇单甲醚-聚乳酸嵌段共聚物(mPEG-PDLLA)作为药物载体,通过溶液吸收法,用旋转蒸发仪蒸发溶剂后制得吲哚美辛固化聚合物胶束(IND-SPM).取IND 20 mg,以PBs 250 mL为溶出介质进行体外溶出度实验.结果表明,30 min内吲哚美辛的释放量高于90%(w/w).DSC,1H NMR和SEM结果证明IND被包裹在mPEG-PDLLA中.随着聚合物的用量的增加,吲哚美辛的溶解度可提高4.6倍.IND-SPM适用于开发成片剂或胶囊剂.%In this study, indomethacin (IND) loaded solidified-polymeric micelles (IND-SPM) were prepared.Their in vitro characteristics were investigated. Methoxy-poly(ethylene glycol) poly(D, L-lactide) copolymer (mPEG-PDLLA) was used as IND carrier. The preparation of IND-SPM was conducted by solution-absorption method and evaporation by rotary evaporator. Polyplasdone XL-10 was used as adsorbent. The solution-absorption method was conducted by the following procedure; IND and mPEG-PDLLA were dissolved in acetone, followed by addition of polyplasdone XL-10 and stirred to obtain a suspension. The powder of IND-SPM was simply obtained after the organic solvent was completely evaporated. More than 90% (w/w) of IND (20 mg) in the powder was dissolved in 250 mL PBS within 30 min. DSC, 1H NMR and SEM results proved that IND was encapsulated within mPEG-PDLLA. The solubility of IND in the system increased 4.6 times with the highest amount of copolymer. The solidified particles were found to be suitable for the formulation of tablets or capsules.

  4. Gemcitabine-loaded smart carbon nanotubes for effective targeting to cancer cells.

    Science.gov (United States)

    Singh, Ravendra; Mehra, Neelesh Kumar; Jain, Vikas; Jain, Narendra Kumar

    2013-07-01

    Carbon nanotubes (CNTs) are the three-dimensional sp(2) hybridized nano-containers that have attracted considerable interest in drug delivery by offering potential advantages such as biocompatibility, non-immunogenicity, high loading efficiency, intrinsic stability and low toxicities. The aim of the present investigation was to assess the potential of gemcitabine-loaded folic acid (FA) conjugated multi-walled CNTs (GEM/FA-NT) for targeting to breast cancer cells. Pristine MWCNTs was functionalized by FA followed by carboxylation, acylation and amidation and characterized by electron microscopy, FT-IR spectroscopy, X-ray diffraction, entrapment efficiency, cytotoxicity and in vivo studies. FDA-approved GEM was loaded to the purified (GEM-NT) and GEM/FA-NT, and % entrapment efficiency was found to be approximately 71.60 ± 0.25 and 79.60 ± 0.45, respectively. The developed formulation GEM/FA-NT was found to have significantly less hemolytic toxicity (8.23 ± 0.65) as compared to free GEM (17.34 ± 0.56). The in vitro release was found to be in sustained pattern at the lysosomal pH, which depicts more cytotoxic response on human breast cancer cell line (MCF-7). It may be interpreted that the GEM/FA-NT formulation is capable to carry drug and deliver it selectively at the tumor site while minimizing side effects and thus holds promise in chemotherapy.

  5. Gemcitabine-loaded albumin nanospheres (GEM-ANPs) inhibit PANC-1 cells in vitro and in vivo

    Science.gov (United States)

    Li, Ji; Di, Yang; Jin, Chen; Fu, Deliang; Yang, Feng; Jiang, Yongjian; Yao, Lie; Hao, Sijie; Wang, Xiaoyi; Subedi, Sabin; Ni, Quanxing

    2013-04-01

    With the development of nanotechnology, special attention has been given to the nanomaterial application in tumor treatment. Here, a modified desolvation-cross-linking method was successfully applied to fabricate gemcitabine-loaded albumin nanospheres (GEM-ANPs), with 110 and 406 nm of mean diameter, respectively. The aim of this study was to assess the drug distribution, side effects, and antitumor activity of GEM-ANPs in vivo. The metabolic viability and flow cytometry analysis revealed that both GEM-ANPs, especially 406-nm GEM-ANPs, could effectively inhibit the metabolism and proliferation and promote the apoptosis of human pancreatic carcinoma (PANC-1) in vitro. Intravenous injection of 406-nm GEM-ANPs exhibited a significant increase of gemcitabine in the pancreas, liver, and spleen of Sprague-Dawley rats ( p tumor mice, intravenous injection of 406-nm GEM-ANPs also could effectively reduce the tumor volume by comparison with free gemcitabine. With these findings, albumin nanosphere-loading approach might be efficacious to improve the antitumor activity of gemcitabine, and the efficacy is associated with the size of GEM-ANPs.

  6. Physicochemical Biomolecular Insights into Buffalo Milk-Derived Nanovesicles.

    Science.gov (United States)

    Baddela, Vijay Simha; Nayan, Varij; Rani, Payal; Onteru, Suneel Kumar; Singh, Dheer

    2016-02-01

    Milk is a natural nutraceutical produced by mammals. The nanovesicles of milk play a role in horizontal gene transfer and confer health-benefits to milk consumers. These nanovesicles contain miRNA, mRNA, and proteins which mediate the intercellular communication. In this work, we isolated and characterized the buffalo milk-derived nanovesicles by dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), scanning electron microscopy (SEM), Western probing, and Fourier transform infrared (FTIR) spectroscopy. The DLS data suggested a bimodal size distribution with one mode near 50 nm and the other around 200 nm for the nanovesicles. The NTA and SEM data also supported the size of nanovesicles within a range of 50-200 nm. The FTIR measurements of nanovesicles identified some prominent absorption bands attributable to the proteins (1300-1700 cm(-1), amide A and amide B bands), lipids (2800-3100 cm(-1)), polysaccharides, and nucleic acids (900-1200 cm(-1)). The comparative expression profiles of immune miRNA signatures (miR-15b, miR-21, miR-27b, miR-125b, miR-155, and miR-500) in nanovesicles isolated from milk, serum, and urine revealed that these miRNAs are present abundantly (P compounds from buffalo milk with high proportion of stable immune miRNAs compared to urine and plasma of same animals.

  7. Large-scale generation of cell-derived nanovesicles

    Science.gov (United States)

    Jo, W.; Kim, J.; Yoon, J.; Jeong, D.; Cho, S.; Jeong, H.; Yoon, Y. J.; Kim, S. C.; Gho, Y. S.; Park, J.

    2014-09-01

    Exosomes are enclosed compartments that are released from cells and that can transport biological contents for the purpose of intercellular communications. Research into exosomes is hindered by their rarity. In this article, we introduce a device that uses centrifugal force and a filter with micro-sized pores to generate a large quantity of cell-derived nanovesicles. The device has a simple polycarbonate structure to hold the filter, and operates in a common centrifuge. Nanovesicles are similar in size and membrane structure to exosomes. Nanovesicles contain intracellular RNAs ranging from microRNA to mRNA, intracellular proteins, and plasma membrane proteins. The quantity of nanovesicles produced using the device is 250 times the quantity of naturally secreted exosomes. Also, the quantity of intracellular contents in nanovesicles is twice that in exosomes. Nanovesicles generated from murine embryonic stem cells can transfer RNAs to target cells. Therefore, this novel device and the nanovesicles that it generates are expected to be used in exosome-related research, and can be applied in various applications such as drug delivery and cell-based therapy.

  8. Adipose stem cell-derived nanovesicles inhibit emphysema primarily via an FGF2-dependent pathway.

    Science.gov (United States)

    Kim, You-Sun; Kim, Ji-Young; Cho, RyeonJin; Shin, Dong-Myung; Lee, Sei Won; Oh, Yeon-Mok

    2017-01-13

    Cell therapy using stem cells has produced therapeutic benefits in animal models of COPD. Secretory mediators are proposed as one mechanism for stem cell effects because very few stem cells engraft after injection into recipient animals. Recently, nanovesicles that overcome the disadvantages of natural exosomes have been generated artificially from cells. We generated artificial nanovesicles from adipose-derived stem cells (ASCs) using sequential penetration through polycarbonate membranes. ASC-derived artificial nanovesicles displayed a 100 nm-sized spherical shape similar to ASC-derived natural exosomes and expressed both exosomal and stem cell markers. The proliferation rate of lung epithelial cells was increased in cells treated with ASC-derived artificial nanovesicles compared with cells treated with ASC-derived natural exosomes. The lower dose of ASC-derived artificial nanovesicles had similar regenerative capacity compared with a higher dose of ASCs and ASC-derived natural exosomes. In addition, FGF2 levels in the lungs of mice treated with ASC-derived artificial nanovesicles were increased. The uptake of ASC-derived artificial nanovesicles was inhibited by heparin, which is a competitive inhibitor of heparan sulfate proteoglycan that is associated with FGF2 signaling. Taken together, the data indicate that lower doses of ASC-derived artificial nanovesicles may have beneficial effects similar to higher doses of ASCs or ASC-derived natural exosomes in an animal model with emphysema, suggesting that artificial nanovesicles may have economic advantages that warrant future clinical studies.

  9. Effects of the composite nanovesicles on the physical properties and cellular adhesion of chitosan films.

    Science.gov (United States)

    Lionzo, Maria I Z; Lorenzini, Giulia C; Tomedi, Joelson; Pranke, Patricia; Silveira, Nádya P

    2012-04-01

    Chitosan films were prepared by the casting of a chitosan gel in absence and presence of composite nanovesicles. The microscopy images showed the occurrence of agglomerates on the surface and internal pores when the nanovesicles were added to the films, differently from the smooth surface of the pure chitosan films. Despite the hydrophobic character that composite nanovesicles gave to the chitosan films, as showed by the reduction of the water permeation at prolonged times, there was a reduction on the contact angle values for these samples related to the roughness of the surface. The peak of water desorption observed on calorimetric analysis of chitosan was shifted to higher values when the nanovesicles were added to the films. Furthermore, the desappearance of Tg on the films containing nanovesicles denoted their plastifier effect in the chitosan film. The swelling results showed higher water diffusion at the first times for the films containing nanovesicles because of the pores observed by microscopy. However, at prolonged times, there was a reduction on the swelling because of the lipofilic composition of the nanovesicles. Furthermore, the presence of nanovesicles led to a reduction on the water content in the chitosan films. Due to the effect on the physical properties of the chitosan films, the addition of nanovesicles on discrete concentrations contributed to the cell adhesion.

  10. Adipose stem cell-derived nanovesicles inhibit emphysema primarily via an FGF2-dependent pathway

    Science.gov (United States)

    Kim, You-Sun; Kim, Ji-Young; Cho, RyeonJin; Shin, Dong-Myung; Lee, Sei Won; Oh, Yeon-Mok

    2017-01-01

    Cell therapy using stem cells has produced therapeutic benefits in animal models of COPD. Secretory mediators are proposed as one mechanism for stem cell effects because very few stem cells engraft after injection into recipient animals. Recently, nanovesicles that overcome the disadvantages of natural exosomes have been generated artificially from cells. We generated artificial nanovesicles from adipose-derived stem cells (ASCs) using sequential penetration through polycarbonate membranes. ASC-derived artificial nanovesicles displayed a 100 nm-sized spherical shape similar to ASC-derived natural exosomes and expressed both exosomal and stem cell markers. The proliferation rate of lung epithelial cells was increased in cells treated with ASC-derived artificial nanovesicles compared with cells treated with ASC-derived natural exosomes. The lower dose of ASC-derived artificial nanovesicles had similar regenerative capacity compared with a higher dose of ASCs and ASC-derived natural exosomes. In addition, FGF2 levels in the lungs of mice treated with ASC-derived artificial nanovesicles were increased. The uptake of ASC-derived artificial nanovesicles was inhibited by heparin, which is a competitive inhibitor of heparan sulfate proteoglycan that is associated with FGF2 signaling. Taken together, the data indicate that lower doses of ASC-derived artificial nanovesicles may have beneficial effects similar to higher doses of ASCs or ASC-derived natural exosomes in an animal model with emphysema, suggesting that artificial nanovesicles may have economic advantages that warrant future clinical studies. PMID:28082743

  11. Erythrocyte-derived optical nano-vesicles as theranostic agents

    Science.gov (United States)

    Mac, Jenny T.; Nunez, Vicente; Bahmani, Baharak; Guerrero, Yadir; Tang, Jack; Vullev, Valentine I.; Anvari, Bahman

    2015-07-01

    We have engineered nano-vesicles, derived from erythrocytes, which can be doped with various near infrared (NIR) organic chromophores, including the FDA-approved indocyanine green (ICG). We refer to these vesicles as NIR erythrocyte-mimicking transducers (NETS) since in response to NIR photo-excitation they can generate heat or emit fluorescent light. Using biochemical methods based on reduction amination, we have functionalized the surface of NET with antibodies to target specific biomolecules. We present results that demonstrate the effectiveness of NETs in targeted imaging of cancer cells that over-express the human epidermal growth factor receptor-2 (HER2).

  12. Hollow Alveolus-Like Nanovesicle Assembly with Metal-Encapsulated Hollow Zeolite Nanocrystals.

    Science.gov (United States)

    Dai, Chengyi; Zhang, Anfeng; Liu, Min; Gu, Lin; Guo, Xinwen; Song, Chunshan

    2016-08-23

    Inspired by the vesicular structure of alveolus which has a porous nanovesicle structure facilitating the transport of oxygen and carbon dioxide, we designed a hollow nanovesicle assembly with metal-encapsulated hollow zeolite that would enhance diffusion of reactants/products and inhibit sintering and leaching of active metals. This zeolitic nanovesicle has been successfully synthesized by a strategy which involves a one-pot hydrothermal synthesis of hollow assembly of metal-containing solid zeolite crystals without a structural template and a selective desilication-recrystallization accompanied by leaching-hydrolysis to convert the metal-containing solid crystals into metal-encapsulated hollow crystals. We demonstrate the strategy in synthesizing a hollow nanovesicle assembly of Fe2O3-encapsulated hollow crystals of ZSM-5 zeolite. This material possesses a microporous (0.4-0.6 nm) wall of hollow crystals and a mesoporous (5-17 nm) shell of nanovesicle with macropores (about 350 nm) in the core. This hierarchical structure enables excellent Fe2O3 dispersion (3-4 nm) and resistance to sintering even at 800 °C; facilitates the transport of reactant/products; and exhibits superior activity and resistance to leaching in phenol degradation. Hollow nanovesicle assembly of Fe-Pt bimetal-encapsulated hollow ZSM-5 crystals was also prepared.

  13. Synthesis and nonlinear optical response of silver nanoparticles decorated polydiacetylene composite nanovesicles

    Science.gov (United States)

    Bhushan, B.; Kundu, T.; Singh, B. P.

    2014-02-01

    We have synthesized, characterized and studied the third-order nonlinear optical properties of polydiacetylene (PDA) nanovesicles decorated by silver nanoparticles. The second molecular hyperpolarizability γ (- ω ; ω , - ω , ω) of the sample was investigated by the antiresonant ring interferometric nonlinear spectroscopic (ARINS) technique using femtosecond modelocked Ti:sapphire laser in the spectral range of 720-820 nm. The observed dispersion of γ has been explained in the framework of three-essential states model involving the ground state, a one-photon excited state and a two-photon excited state. The energy of two-photon state, transition dipole moments and line width of the transitions have been estimated. Our investigation reveals that the spectral dispersion characteristic of γ for coated PDA nanovesicles is qualitatively similar to that observed for uncoated PDA nanovesicles but bears no resemblance to that observed in silver nanoparticles. The presence of silver nanoparticles increases the γ values of coated nanovesicles slightly as compared to that of uncoated nanovesicles, suggesting a definite but weak coupling between free electrons of metal nanoparticles and π -electrons of polymer in the composite system.

  14. Living nanovesicles--chemical and physical survival strategies of primordial biosystems.

    Science.gov (United States)

    Sommer, Andrei P; McKay, David S; Ciftcioglu, Neva; Oron, Uri; Mester, Adam R; Kajander, E Olavi

    2003-01-01

    Life on Earth and Mars could have started with self-assembled nanovesicles similar to the present nanobacteria (NB). To resist extreme environmental stress situations and periods of nutritional deprivation, nanovesicles would have had a chemical composition protected by a closed mineralized compartment, facilitating their development in a primordial soup, or other early wet environment. Their survivability would have been enhanced if they had mechanisms for metabolic communication, and an ability to collect primordially available energy forms. Here, we establish an irreducible model system for life formation starting with NB.

  15. 紫杉烷类 PEG-PDLLA 纳米粒的制备及其体外评价%Preparation and in-vitro evaluation of PEG-PDLLA in nanoparticle containing taxanes

    Institute of Scientific and Technical Information of China (English)

    魏炜; 张英新; 张晓敏; 蒋成君; 周一峰

    2015-01-01

    采用乳化‐溶剂蒸发法制备紫杉烷类 PEG‐PDLLA纳米粒,马尔文激光粒度仪测其粒径及Zeta电位;HPLC法测定纳米粒包封率和载药量;研究载药纳米粒在 PBS 中的释放动力学;初步评价载药纳米粒在MGC803、HeLa细胞中的摄取及细胞毒性。结果表明,通过包载形成的纳米粒的粒径为(13±1)nm ,分布较集中。载体与药物的质量比在20∶1时,紫杉醇的均一性最好,卡巴他赛的包封率最高,达到88.77%。载药纳米粒具有较好的缓释作用,MGC803、HeLa细胞的存活率降低,与临床用注射剂效果相近。紫杉烷类 PEG‐PDLLA纳米粒的性质、释放、细胞抑瘤率都较好,可为开发紫杉烷类新型静脉注射制剂提供实验依据。%PEG‐PDLLA in nanoparticle containing taxanes was prepared by emulsion solvent evaporation method .The particle size and Zeta potential were evaluated by Malvern Zerta sizer . The encapsulation efficiency and drug‐loading content were examined by HPLC method .The kinetics of taxanes released from the nanoparticle in phosphate buffer solution (PBS ) was studied .The uptake and cell toxicity of drug‐loading nanoparticle in MGC803 and HeLa were preliminary evaluated .The result showed that the particle size is (13 ± 1) nm ,and distribution is concentrated .T he ratio of the carrier and the drug is preferably 20∶1 .T he homogeneity of taxol is best , and encapsulation efficiency of cabazitaxel is highest , reach 88 .77% . T he nanoparticle with drug has a better sustained release .Survival of cancer cells in vitro anti‐tumor decreased with similar clinical results with injections .The nature ,release ,and cell inhibition rate of PEG‐PDLLA in nanoparticle containing taxanes is good .The test provides an experimental basis for the development of novel taxanes intravenous formulations .

  16. Sunitinib microspheres based on [PDLLA-PEG-PDLLA]-b-PLLA multi-block copolymers for ocular drug delivery.

    Science.gov (United States)

    Ramazani, F; Hiemstra, C; Steendam, R; Kazazi-Hyseni, F; Van Nostrum, C F; Storm, G; Kiessling, F; Lammers, T; Hennink, W E; Kok, R J

    2015-09-01

    Sunitinib is a multi-targeted receptor tyrosine kinase (RTK) inhibitor that blocks several angiogenesis related pathways. The aim of this study was to develop sunitinib-loaded polymeric microspheres that can be used as intravitreal formulation for the treatment of ocular diseases. A series of novel multi-block copolymers composed of amorphous blocks of poly-(D,L-lactide) (PDLLA) and polyethylene glycol (PEG) and of semi-crystalline poly-(L-lactide) (PLLA) blocks were synthesized. Sunitinib-loaded microspheres were prepared by a single emulsion method using dichloromethane as volatile solvent and DMSO as co-solvent. SEM images showed that the prepared microspheres (∼ 30 μm) were spherical with a non-porous surface. Sunitinib-loaded microspheres were studied for their degradation and in-vitro release behavior. It was found that increasing the percentage of amorphous soft blocks from 10% to 30% accelerated the degradation of the multi-block copolymers. Sunitinib microspheres released their cargo for a period of at least 210 days by a combination of diffusion and polymer erosion. The initial burst (release in 24h) and release rate could be tailored by controlling the PEG-content of the multi-block copolymers. Sunitinib-loaded microspheres suppressed angiogenesis in a chicken chorioallantoic membrane (CAM) assay. These microspheres therefore hold promise for long-term suppression of ocular neovascularization.

  17. Sunitinib microspheres based on [PDLLA-PEG-PDLLA]-b-PLLA multi-block copolymers for ocular drug delivery

    NARCIS (Netherlands)

    Ramazani, F.; Hiemstra, C.; Steendam, R.; Kazazi-Hyseni, F.; Van Nostrum, C. F.; Storm, G.; Kiessling, F.; Lammers, T.; Hennink, W. E.; Kok, R. J.

    2015-01-01

    Sunitinib is a multi-targeted receptor tyrosine kinase (RTK) inhibitor that blocks several angiogenesis related pathways. The aim of this study was to develop sunitinib-loaded polymeric microspheres that can be used as intravitreal formulation for the treatment of ocular diseases. A series of novel

  18. Embryonic cerebrospinal fluid nanovesicles carry evolutionarily conserved molecules and promote neural stem cell amplification.

    Directory of Open Access Journals (Sweden)

    David M Feliciano

    Full Text Available During brain development, neural stem cells (NSCs receive on-or-off signals important for regulating their amplification and reaching adequate neuron density. However, how a coordinated regulation of intracellular pathways and genetic programs is achieved has remained elusive. Here, we found that the embryonic (e CSF contains 10¹² nanoparticles/ml (77 nm diameter, some of which were identified as exosome nanovesicles that contain evolutionarily conserved molecules important for coordinating intracellular pathways. eCSF nanovesicles collected from rodent and human embryos encapsulate protein and microRNA components of the insulin-like growth factor (IGF signaling pathway. Supplementation of eCSF nanovesicles to a mixed culture containing eNSCs activated the IGF-mammalian target of rapamycin complex 1 (mTORC1 pathway in eNSCs and expanded the pool of proliferative eNSCs. These data show that the eCSF serves as a medium for the distribution of nanovesicles, including exosomes, and the coordinated transfer of evolutionary conserved molecules that regulate eNSC amplification during corticogenesis.

  19. Targeting and cytotoxicity of SapC-DOPS nanovesicles in pancreatic cancer.

    Directory of Open Access Journals (Sweden)

    Zhengtao Chu

    Full Text Available Only a small number of promising drugs target pancreatic cancer, which is the fourth leading cause of cancer deaths with a 5-year survival of less than 5%. Our goal is to develop a new biotherapeutic agent in which a lysosomal protein (saposin C, SapC and a phospholipid (dioleoylphosphatidylserine, DOPS are assembled into nanovesicles (SapC-DOPS for treating pancreatic cancer. A distinguishing feature of SapC-DOPS nanovesicles is their high affinity for phosphatidylserine (PS rich microdomains, which are abnormally exposed on the membrane surface of human pancreatic tumor cells. To evaluate the role of external cell PS, in vitro assays were used to correlate PS exposure and the cytotoxic effect of SapC-DOPS in human tumor and nontumorigenic pancreatic cells. Next, pancreatic tumor xenografts (orthotopic and subcutaneous models were used for tumor targeting and therapeutic efficacy studies with systemic SapC-DOPS treatment. We observed that the nanovesicles selectively killed human pancreatic cancer cells in vitro by inducing apoptotic death, whereas untransformed cells remained unaffected. This in vitro cytotoxic effect correlated to the surface exposure level of PS on the tumor cells. Using xenografts, animals treated with SapC-DOPS showed clear survival benefits and their tumors shrank or disappeared. Furthermore, using a double-tracking method in live mice, we showed that the nanovesicles were specifically targeted to orthotopically-implanted, bioluminescent pancreatic tumors. These data suggest that the acidic phospholipid PS is a biomarker for pancreatic cancer that can be effectively targeted for therapy utilizing cancer-selective SapC-DOPS nanovesicles. This study provides convincing evidence in support of developing a new therapeutic approach to pancreatic cancer.

  20. Host-guest interaction induced supramolecular amphiphilic star architecture and uniform nanovesicle formation for anticancer drug delivery.

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    Zhu, Jing-Ling; Liu, Kerh Li; Wen, Yuting; Song, Xia; Li, Jun

    2016-01-21

    A star polymer of poly[(R,S)-3-hydroxybutyrate] (PHB) with adamantyl end-terminals extended from an α-cyclodextrin (α-CD) core is designed. It subsequently self-assembles to form controllable and uniform nanovesicles induced by host-guest interactions between heptakis(2,6-di-O-methyl)-β-CD and the adamantyl ends. The nanovesicles are suitable for loading and intracellular delivery of the anticancer drug doxorubicin.

  1. Phosphatidylcholine nanovesicles coated with chitosan or chondroitin sulfate as novel devices for bacteriocin delivery

    Science.gov (United States)

    da Silva, Indjara Mallmann; Boelter, Juliana Ferreira; da Silveira, Nádya Pesce; Brandelli, Adriano

    2014-07-01

    There is increased interest on the use of natural antimicrobial peptides in biomedicine and food preservation technologies. In this study, the antimicrobial activity of nisin encapsulated into nanovesicles containing polyanionic polysaccharides was investigated. Nisin was encapsulated in phosphatidylcholine (PC) liposomes containing chitosan or chondroitin sulfate by the thin-film hydration method and tested for antimicrobial activity against Listeria spp. The mean particle size of PC liposomes was 145 nm and varied to 210 and 134 nm with the incorporation of chitosan and chondroitin sulfate, respectively. Nisin-containing nanovesicles with and without incorporation of polysaccharides had a zeta potential values around -20 mV, showing mostly spherical structures when observed by transmission electron microscopy. Encapsulated nisin had similar efficiency as free nisin in inhibiting Listeria spp. isolated from bovine carcass, and greater efficiency in inhibiting Listeria monocytogenes. The formulation containing chitosan was more stable and more efficient in inhibiting L. monocytogenes when compared to the other nanovesicles tested. After 24 h, the viable cell counts were 2 log lower as compared with the other treatments and 7 log comparing to controls.

  2. Dual stimuli polysaccharide nanovesicles for conjugated and physically loaded doxorubicin delivery in breast cancer cells

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    Pramod, P. S.; Shah, Ruchira; Jayakannan, Manickam

    2015-04-01

    The present work reports the development of pH and enzyme dual responsive polysaccharide vesicular nano-scaffolds for the administration of doxorubicin via physical loading and polymer-drug conjugation to breast cancer cells. Dextran was suitably modified with a renewable resource 3-pentadecyl phenol unit through imine and aliphatic ester chemical linkages that acted as pH and esterase enzyme stimuli, respectively. These dual responsive polysaccharide derivatives self-organized into 200 +/- 10 nm diameter nano-vesicles in water. The water soluble anticancer drug doxorubicin (DOX.HCl) was encapsulated in the hydrophilic pocket to produce core-loaded polysaccharide vesicles whereas chemical conjugation produced DOX anchored at the hydrophobic layer of the dextran nano-vesicles. In vitro studies revealed that about 70-80% of the drug was retained under circulatory conditions at pH = 7.4 and 37 °C. At a low pH of 6.0 to 5.0 and in the presence of esterase; both imine and ester linkages were cleaved instantaneously to release 100% of the loaded drugs. Cytotoxicity assays on Wild Type Mouse Embryonic Fibroblasts (WTMEFs) confirmed the non-toxicity of the newly developed dextran derivatives at up to 500 μg mL-1 in PBS. MTT assays on fibroblast cells revealed that DOX.HCl loaded nano-vesicles exhibited better killing abilities than DOX conjugated polymer nano-vesicles. Both DOX loaded and DOX conjugated nano-vesicles were found to show significant killing in breast cancer cells (MCF 7). Confocal microscopy images confirmed the uptake of DOX loaded (or conjugated) nano-vesicles by cells compared to free DOX. Thus, the newly developed pH and enzyme dual responsive polysaccharide vesicular assemblies are potential drug vectors for the administration of DOX in both loaded and chemically conjugated forms for the efficient killing of breast cancer cells.The present work reports the development of pH and enzyme dual responsive polysaccharide vesicular nano-scaffolds for the

  3. Therapeutic effects of autologous tumor-derived nanovesicles on melanoma growth and metastasis.

    Directory of Open Access Journals (Sweden)

    Eun-Young Lee

    Full Text Available Cancer vaccines with optimal tumor-associated antigens show promise for anti-tumor immunotherapy. Recently, nano-sized vesicles, such as exosomes derived from tumors, were suggested as potential antigen candidates, although the total yield of exosomes is not sufficient for clinical applications. In the present study, we developed a new vaccine strategy based on nano-sized vesicles derived from primary autologous tumors. Through homogenization and sonication of tumor tissues, we achieved high yields of vesicle-bound antigens. These nanovesicles were enriched with antigenic membrane targets but lacked nuclear autoantigens. Furthermore, these nanovesicles together with adjuvant activated dendritic cells in vitro, and induced effective anti-tumor immune responses in both primary and metastatic melanoma mouse models. Therefore, autologous tumor-derived nanovesicles may represent a novel source of antigens with high-level immunogenicity for use in acellular vaccines without compromising safety. Our strategy is cost-effective and can be applied to patient-specific cancer therapeutic vaccination.

  4. Force measurements on natural membrane nanovesicles reveal a composition-independent, high Young's modulus

    Science.gov (United States)

    Calò, Annalisa; Reguera, David; Oncins, Gerard; Persuy, Marie-Annick; Sanz, Guenhaël; Lobasso, Simona; Corcelli, Angela; Pajot-Augy, Edith; Gomila, Gabriel

    2014-01-01

    Mechanical properties of nano-sized vesicles made up of natural membranes are crucial to the development of stable, biocompatible nanocontainers with enhanced functional, recognition and sensing capabilities. Here we measure and compare the mechanical properties of plasma and inner membrane nanovesicles ~80 nm in diameter obtained from disrupted yeast Saccharomyces cerevisiae cells. We provide evidence of a highly deformable behaviour for these vesicles, able to support repeated wall-to-wall compressions without irreversible deformations, accompanied by a noticeably high Young's modulus (~300 MPa) compared to that obtained for reconstituted artificial liposomes of similar size and approaching that of some virus particles. Surprisingly enough, the results are approximately similar for plasma and inner membrane nanovesicles, in spite of their different lipid compositions, especially on what concerns the ergosterol content. These results point towards an important structural role of membrane proteins in the mechanical response of natural membrane vesicles and open the perspective to their potential use as robust nanocontainers for bioapplications.Mechanical properties of nano-sized vesicles made up of natural membranes are crucial to the development of stable, biocompatible nanocontainers with enhanced functional, recognition and sensing capabilities. Here we measure and compare the mechanical properties of plasma and inner membrane nanovesicles ~80 nm in diameter obtained from disrupted yeast Saccharomyces cerevisiae cells. We provide evidence of a highly deformable behaviour for these vesicles, able to support repeated wall-to-wall compressions without irreversible deformations, accompanied by a noticeably high Young's modulus (~300 MPa) compared to that obtained for reconstituted artificial liposomes of similar size and approaching that of some virus particles. Surprisingly enough, the results are approximately similar for plasma and inner membrane nanovesicles, in

  5. ACECLOFENAC ENCAPSULATED ETHANOLIC NANO-VESICLES FOR EFFECTIVE TREATMENT OF OSTEOART HRITIS

    Directory of Open Access Journals (Sweden)

    Arvinder Kaur et al

    2012-08-01

    Full Text Available In the present study, ethanolic nanovesicles of Aceclofenac developed for the site specific delivery to joints for effective treatment of osteoarthritis. Ethanolic nano-vesicles were prepared by solvent dispersion method. Vesicles were characterized for vesicular size, surface morphology, size and size distribution, zeta potential, entrapment efficiency. Formulations were also evaluated for drug-vesicle (excipients interaction, in vitro permeation, in vitro deposition. The TEM showed dark vesicular structures by which it is possible to notice the outer most bilayer. In the ethanol concentration range of 20-40%, the size of the vesicles increased with decreasing ethanol concentration, with the largest vesicles in preparations containing 20% ethanol (809.34±2.329 nm and the smallest in preparations containing 40% ethanol (627±3.684 nm. Zeta potential of formulations shows negative value, which indicates that, the higher concentration of ethanol make the negative charge on vesicles surface. The entrapment was found to increase with increase in ethanol concentration, however percent entrapment decreased when ethanol concentration exceeded 40%. The FTIR spectra reveled no considerable change in IR peaks of Aceclofenac loaded ethanolic vesicles when compared to pure drugs there by indicating absence of any interaction.

  6. Competition between Bending and Internal Pressure Governs the Mechanics of Fluid Nanovesicles.

    Science.gov (United States)

    Vorselen, Daan; MacKintosh, Fred C; Roos, Wouter H; Wuite, Gijs J L

    2017-03-28

    Nanovesicles (∼100 nm) are ubiquitous in cell biology and an important vector for drug delivery. Mechanical properties of vesicles are known to influence cellular uptake, but the mechanism by which deformation dynamics affect internalization is poorly understood. This is partly due to the fact that experimental studies of the mechanics of such vesicles remain challenging, particularly at the nanometer scale where appropriate theoretical models have also been lacking. Here, we probe the mechanical properties of nanoscale liposomes using atomic force microscopy (AFM) indentation. The mechanical response of the nanovesicles shows initial linear behavior and subsequent flattening corresponding to inward tether formation. We derive a quantitative model, including the competing effects of internal pressure and membrane bending, that corresponds well to these experimental observations. Our results are consistent with a bending modulus of the lipid bilayer of ∼14kbT. Surprisingly, we find that vesicle stiffness is pressure dominated for adherent vesicles under physiological conditions. Our experimental method and quantitative theory represents a robust approach to study the mechanics of nanoscale vesicles, which are abundant in biology, as well as being of interest for the rational design of liposomal vectors for drug delivery.

  7. Bioelectronic Device Mimicking Human Sensory System based on Nanovesicle-Carbon Nanotube Hybrid Structure

    Science.gov (United States)

    Kim, Daesan; Jin, Hye; Lee, San; Kim, Tae; Park, Juhun; Song, Hyun; Park, Tai; Hong, Seunghun

    2013-03-01

    We have developed a nanovesicle-based bioelectronic nose (NBN) that could mimic the receptor-mediated signal transmission of human olfactory systems and recognize a specific odorant. The NBN was comprised of a single-walled carbon nanotube (CNT)-based field effect transistor and cell-derived nanovesicles containing human olfactory receptors and calcium ion signal pathways. Importantly, the NBN took advantages of cell signal pathways for sensing signal amplification. It enabled ~100 times higher sensitivity than that of previous bioelectronic noses based on only olfactory receptor protein and CNT transistors. The NBN sensors exhibited a high sensitivity of 1 fM detection limit and a human-like selectivity with single-carbon-atomic resolution. Furthermore, these sensors could mimic a receptor-mediated cellular signal transmission in live cells. This versatile sensor platform should be useful for the study of molecular recognition and biological processes on cell membranes and also for various practical applications such as food conditioning and medical diagnostics.

  8. pH-Responsive reversible self-assembly of gold nanoparticles into nanovesicles

    Science.gov (United States)

    Fan, Chunfang; Bian, Tong; Shang, Lu; Shi, Run; Wu, Li-Zhu; Tung, Chen-Ho; Zhang, Tierui

    2016-02-01

    A novel system was reported to realize the reversible self-assembly and disassembly of Au nanovesicles (NVs) driven by pH stimuli with commercially available organic molecules, 4-mercaptobenzonic acid (4-MBA) and oleylamine (OL). Through adjusting deprotonation and protonation of 4-MBA, Au NVs demonstrated a good reversible self-assembly behavior. As a proof-of-concept, Rhodamine B was loaded into the vesicles to demonstrate the reversible pH-responsive controlled release.A novel system was reported to realize the reversible self-assembly and disassembly of Au nanovesicles (NVs) driven by pH stimuli with commercially available organic molecules, 4-mercaptobenzonic acid (4-MBA) and oleylamine (OL). Through adjusting deprotonation and protonation of 4-MBA, Au NVs demonstrated a good reversible self-assembly behavior. As a proof-of-concept, Rhodamine B was loaded into the vesicles to demonstrate the reversible pH-responsive controlled release. Electronic supplementary information (ESI) available: Experimental details, characterization, and Fig. S1-S4. See DOI: 10.1039/c6nr00044d

  9. Micro- and Nano-vesicles from First Trimester Human Placentae Carry Flt-1 and Levels Are Increased in Severe Preeclampsia.

    Science.gov (United States)

    Tong, Mancy; Chen, Qi; James, Joanna L; Stone, Peter R; Chamley, Lawrence W

    2017-01-01

    Preeclampsia is a life-threatening hypertensive disease affecting 3-5% of pregnancies. While the pathogenesis of preeclampsia remains unclear, it is known that placenta-derived factors trigger the disease by activating maternal endothelial cells prior to the onset of clinical symptoms. Extracellular vesicles (EVs) of different sizes extruded by the placenta may be one factor. The truncated/secreted form of Flt-1 (sFlt-1) has also been implicated in the pathogenesis of preeclampsia. We investigated whether placental EV production is altered in preeclampsia such that they induce endothelial cell activation, and whether (s)Flt-1 is involved. Macro-, micro-, and nano-vesicles were collected from normal and preeclamptic (PE) placental explants, and separated by differential centrifugation. The number and size of micro- and nano-vesicles was measured by nanoparticle tracking analysis and their ability to activate endothelial cells was quantified by endothelial cell intercellular adhesion molecule 1 expression and monocyte adhesion. The levels of Flt-1 were measured by western blots and ELISA. PE placentae extruded significantly more micro- and nano-vesicles than control placentae and the extruded micro-vesicles were larger than those from control placentae. Micro- and nano-vesicles from both first trimester and term human placentae carried Flt-1 and levels were significantly increased in EVs from severe, but not mild, PE compared to normotensive placentae. All fractions of EVs from PE placentae activated endothelial cells, and for micro- and nano-vesicles, activation was reduced in the presence of exogenous vascular endothelial growth factor (VEGF), a Flt-1 neutralizing antibody, or by pre-treatment with VEGF. While EV-bound VEGF constituted over 20% of the total detected VEGF secreted by PE and normotensive placentae, EV-bound Flt-1 did not significantly contribute to the total level of sFlt-1/Flt-1 released by human third trimester placentae. Micro- and nano-vesicles

  10. Imaging and Therapy of Pancreatic Cancer with Phosphatidylserine-Targeted Nanovesicles1

    Science.gov (United States)

    Blanco, Victor M.; Latif, Tahir; Chu, Zhengtao; Qi, Xiaoyang

    2015-01-01

    Pancreatic cancer remains one of the most intractable cancers, with a dismal prognosis reflected by a 5-year survival of ~ 6%. Since early disease symptoms are undefined and specific biomarkers are lacking, about 80% of patients present with advanced, inoperable tumors that represent a daunting challenge. Despite many clinical trials, no single chemotherapy agent has been reliably associated with objective response rates above 10% or median survival longer than 5 to 7 months. Although combination chemotherapy regimens have in recent years provided some improvement, overall survival (8-11 months) remains very poor. There is therefore a critical need for novel therapies that can improve outcomes for pancreatic cancer patients. Here, we present a summary of the current therapies used in the management of advanced pancreatic cancer and review novel therapeutic strategies that target tumor biomarkers. We also describe our recent research using phosphatidylserine-targeted saposin C–coupled dioleoylphosphatidylserine nanovesicles for imaging and therapy of pancreatic cancer. PMID:26055177

  11. Nanovesicle encapsulation of antimicrobial peptide P34: physicochemical characterization and mode of action on Listeria monocytogenes

    Science.gov (United States)

    da Silva Malheiros, Patrícia; Sant'Anna, Voltaire; Micheletto, Yasmine Miguel Serafini; da Silveira, Nadya Pesce; Brandelli, Adriano

    2011-08-01

    Antimicrobial peptide P34, a substance showing antibacterial activity against pathogenic and food spoilage bacteria, was encapsulated in liposomes prepared from partially purified soybean phosphatidylcholine, and their physicochemical characteristics were evaluated. The antimicrobial activity was estimated by agar diffusion assay using Listeria monocytogenes ATCC 7644 as indicator strain. A concentration of 3,200 AU/mL of P34 was encapsulated in nanovesicles and stocked at 4 °C. No significant difference ( p > 0.05) in the biological activity of free and encapsulated P34 was observed through 24 days. Size and PDI of liposomes, investigated by light scattering analysis, were on average 150 nm and 0.22 respectively. Zeta potential was -27.42 mV. There was no significant change ( p > 0.05) in the physicochemical properties of liposomes during the time of evaluation. The liposomes presented closed spherical morphology as visualized by transmission electron microscopy (TEM). The mode of action of liposome-encapsulated P34 under L. monocytogenes cells was investigated by TEM. Liposomes appeared to adhere but not fuse with the bacterial cell wall, suggesting that the antimicrobial is released from nanovesicles to act against the microorganism. The effect of free and encapsulated P34 was tested against L. monocytogenes, showing that free bacteriocin inhibited the pathogen more quickly than the encapsulated P34. Liposomes prepared with low-cost lipid showed high encapsulation efficiency for a new antimicrobial peptide and were stable during storage. The mode of action against the pathogen L. monocytogenes was characterized.

  12. Endogenous lung surfactant inspired pH responsive nanovesicle aerosols: Pulmonary compatible and site-specific drug delivery in lung metastases

    Science.gov (United States)

    Joshi, Nitin; Shirsath, Nitesh; Singh, Ankur; Joshi, Kalpana S.; Banerjee, Rinti

    2014-11-01

    Concerns related to pulmonary toxicity and non-specificity of nanoparticles have limited their clinical applications for aerosol delivery of chemotherapeutics in lung cancer. We hypothesized that pulmonary surfactant mimetic nanoparticles that offer pH responsive release specifically in tumor may be a possible solution to overcome these issues. We therefore developed lung surfactant mimetic and pH responsive lipid nanovesicles for aerosol delivery of paclitaxel in metastatic lung cancer. 100-200 nm sized nanovesicles showed improved fusogenicity and cytosolic drug release, specifically with cancer cells, thereby resulting in improved cytotoxicity of paclitaxel in B16F10 murine melanoma cells and cytocompatibility with normal lung fibroblasts (MRC 5). The nanovesicles showed airway patency similar to that of endogenous pulmonary surfactant and did not elicit inflammatory response in alveolar macrophages. Their aerosol administration while significantly improving the biodistribution of paclitaxel in comparison to Taxol (i.v.), also showed significantly higher metastastes inhibition (~75%) in comparison to that of i.v. Taxol and i.v. Abraxane. No signs of interstitial pulmonary fiborisis, chronic inflammation and any other pulmonary toxicity were observed with nanovesicle formulation. Overall, these nanovesicles may be a potential platform to efficiently deliver hydrophobic drugs as aerosol in metastatic lung cancer and other lung diseases, without causing pulmonary toxicity.

  13. Effective delivery of recombinant proteins to rod photoreceptors via lipid nanovesicles

    Energy Technology Data Exchange (ETDEWEB)

    Asteriti, Sabrina [Dept. of Translational Research, University of Pisa, Pisa (Italy); Dal Cortivo, Giuditta [Dept. of Life Sciences and Reproduction, University of Verona, Strada Le Grazie 8, Verona (Italy); Pontelli, Valeria [Dept. of Neurological and Movement Sciences, University of Verona, Strada Le Grazie 8, Verona (Italy); Cangiano, Lorenzo [Dept. of Translational Research, University of Pisa, Pisa (Italy); Buffelli, Mario, E-mail: mario.buffelli@univr.it [Dept. of Neurological and Movement Sciences, University of Verona, Strada Le Grazie 8, Verona (Italy); Center for Biomedical Computing, University of Verona, Strada le Grazie 8, 37134 Verona (Italy); Dell’Orco, Daniele, E-mail: daniele.dellorco@univr.it [Dept. of Life Sciences and Reproduction, University of Verona, Strada Le Grazie 8, Verona (Italy); Center for Biomedical Computing, University of Verona, Strada le Grazie 8, 37134 Verona (Italy)

    2015-06-12

    The potential of liposomes to deliver functional proteins in retinal photoreceptors and modulate their physiological response was investigated by two experimental approaches. First, we treated isolated mouse retinas with liposomes encapsulating either recoverin, an important endogenous protein operating in visual phototransduction, or antibodies against recoverin. We then intravitrally injected in vivo liposomes encapsulating either rhodamin B or recoverin and we investigated the distribution in retina sections by confocal microscopy. The content of liposomes was found to be released in higher amount in the photoreceptor layer than in the other regions of the retina and the functional effects of the release were in line with the current model of phototransduction. Our study sets the basis for quantitative investigations aimed at assessing the potential of intraocular protein delivery via biocompatible nanovesicles, with promising implications for the treatment of retinal diseases affecting the photoreceptor layer. - Highlights: • Recombinant proteins encapsulated in nano-sized liposomes injected intravitreally reach retinal photoreceptors. • The phototransduction cascade in rods is modulated by the liposome content. • Mathematical modeling predicts the alteration of the photoresponses following liposome fusion.

  14. Microtubule-associated protein-4 controls nanovesicle dynamics and T cell activation.

    Science.gov (United States)

    Bustos-Morán, Eugenio; Blas-Rus, Noelia; Martin-Cófreces, Noa Beatriz; Sánchez-Madrid, Francisco

    2017-04-01

    The immune synapse (IS) is a specialized structure formed at the contact area between T lymphocytes and antigen-presenting cells (APCs) that is essential for the adaptive immune response. Proper T cell activation requires its polarization towards the APC, which is highly dependent on the tubulin cytoskeleton. Microtubule-associated protein-4 (MAP4) is a microtubule (MT)-stabilizing protein that controls MTs in physiological processes, such as cell division, migration, vesicular transport or primary cilia formation. In this study, we assessed the role of MAP4 in T cell activation. MAP4 decorates the pericentrosomal area and MTs of the T cell, and it is involved in MT detyrosination and stable assembly in response to T cell activation. In addition, MAP4 prompts the timely translocation of the MT-organizing center (MTOC) towards the IS and the dynamics of signaling nanovesicles that sustains T cell activation. However, MAP4 acts as a negative regulator of other T cell activation-related signals, including diacylglycerol (DAG) production and IL2 secretion. Our data indicate that MAP4 acts as a checkpoint molecule that balances positive and negative hallmarks of T cell activation.

  15. Development and characterization of phosphatidylcholine nanovesicles, containing garlic extract, with antilisterial activity in milk.

    Science.gov (United States)

    Pinilla, Cristian Mauricio Barreto; Noreña, Caciano Pelayo Zapata; Brandelli, Adriano

    2017-04-01

    Phospholipid nanovesicles were developed to improve the stability of garlic (Allium sativum L.) extract. Electron microscopy of liposomes revealed nanometric and spherical-shaped vesicles with a mean particle size of 174.6±17.3nm and polydispersity index of 0.26±0.02. The entrapment efficiency was 47.5±7.3% and the nanoliposomes had a zeta potential of -16.2±5.5mV. The antimicrobial activity of free and encapsulated garlic extract was evaluated against different strains of Listeria spp. in milk at 37°C for 24h. For free and encapsulated garlic extracts at 5% concentration, a decrease of 4log cycles in viable cell counts was observed at 10h, against four of the five strains of Listeria spp. tested. The results indicate that liposomes constitute a suitable system for encapsulation of unstable garlic active compounds and the encapsulation of garlic extract proves to be a promising technology for multiple applications, including antimicrobial agents.

  16. Near-infrared spectroscopic studies of self-forming lipids and nanovesicles

    Science.gov (United States)

    Bista, Rajan K.; Bruch, Reinhard F.

    2009-02-01

    Lipids and liposomes have remained an active research topic for several decades due to their significance as membrane model. Several vibrational spectroscopic techniques have been developed and employed to study the properties of lipids and liposomes. In this study, near-infrared (NIR) spectroscopy has been used to analyze a suite of synthesized PEGylated lipids trademarked as QuSomesTM. The three amphiphiles used in this study, differ in their apolar hydrophobic chain length and contain various units of polar polyethylene glycol (PEG) head groups. In contrast to conventional phospholipids, this new kind of lipids forms liposomes spontaneously upon hydration, without the supply of external activation energy. Whilst the NIR spectra of QuSomesTM show a common pattern, differences in the spectra are observed which enable the lipids to be distinguished. NIR absorption spectra of these new artificial lipids have been recorded in the spectral range of 4800-9000 cm-1 (~2100-1100 nm) by using a new miniaturized spectrometer based on micro-optical-electro-mechanical systems (MOEMS) technology. In particular, we have established specific band structures as "molecular fingerprints" corresponding to overtones and combinations vibrational modes involving mainly C-H and O-H functional groups for sample analysis of QuSomesTM. Moreover, we have demonstrated that the nanovesicles formed by such lipids in polar solvents show high stability and obey Beer's law at low concentration. The results reported in this study may find applications in various field including the development of lipids based drug delivery systems.

  17. Citrus limon-derived nanovesicles inhibit cancer cell proliferation and suppress CML xenograft growth by inducing TRAIL-mediated cell death.

    Science.gov (United States)

    Raimondo, Stefania; Naselli, Flores; Fontana, Simona; Monteleone, Francesca; Lo Dico, Alessia; Saieva, Laura; Zito, Giovanni; Flugy, Anna; Manno, Mauro; Di Bella, Maria Antonietta; De Leo, Giacomo; Alessandro, Riccardo

    2015-08-14

    Nanosized vesicles are considered key players in cell to cell communication, thus influencing physiological and pathological processes, including cancer. Nanovesicles have also been found in edible-plants and have shown therapeutic activity in inflammatory bowel diseases; however information on their role in affecting cancer progression is missing.Our study identify for the first time a fraction of vesicles from lemon juice (Citrus limon L.), obtained as a result of different ultracentrifugation, with density ranging from 1,15 to 1,19 g/ml and specific proteomic profile. By using an in vitro approach, we show that isolated nanovesicles inhibit cancer cell proliferation in different tumor cell lines, by activating a TRAIL-mediated apoptotic cell death. Furthermore, we demonstrate that lemon nanovesicles suppress CML tumor growth in vivo by specifically reaching tumor site and by activating TRAIL-mediated apoptotic cell processes. Overall, this study suggests the possible use of plant-edible nanovesicles as a feasible approach in cancer treatment.

  18. Gemcitabine-loaded liposomes: rationale, potentialities and future perspectives

    Directory of Open Access Journals (Sweden)

    Federico C

    2012-11-01

    Full Text Available Cinzia Federico, Valeria M Morittu, Domenico Britti, Elena Trapasso, Donato CoscoDepartment of Health Sciences, Building of BioSciences, University “Magna Græcia” of Catanzaro, Campus Universitario “S Venuta”, Germaneto, ItalyAbstract: This review describes the strategies used in recent years to improve the biopharmaceutical properties of gemcitabine, a nucleoside analog deoxycytidine antimetabolite characterized by activity against many kinds of tumors, by means of liposomal devices. The main limitation of using this active compound is the rapid inactivation of deoxycytidine deaminase following administration in vivo. Consequently, different strategies based on its encapsulation/complexation in innovative vesicular colloidal carriers have been investigated, with interesting results in terms of increased pharmacological activity, plasma half-life, and tumor localization, in addition to decreased side effects. This review focuses on the specific approaches used, based on the encapsulation of gemcitabine in liposomes, with particular attention to the results obtained during the last 5 years. These approaches represent a valid starting point in the attempt to obtain a novel, commercializable drug formulation as already achieved for liposomal doxorubicin (Doxil®, Caelyx®.Keywords: gemcitabine, liposomes, multidrug, poly(ethylene glycol, tumors

  19. Gemcitabine-loaded magnetic albumin nanospheres for cancer chemohyperthermia

    Science.gov (United States)

    Li, Hongbo; Ke, Fei; An, Yanli; Hou, Xinxin; Zhang, Hao; Lin, Mei; Zhang, Dongsheng

    2013-03-01

    Eliminating cancer without harming normal body tissue remains a longstanding challenge in medicine. Toward this goal, we prepared nanosized magnetic albumin nanospheres encapsulating magnetic nanoparticles (Fe3O4) and antitumor drugs (Gemcitabine, GEM). Magnetic albumin nanospheres (average size ≈ 224 nm) had good magnetic responsiveness upon exposure to an alternating magnetic field even though Fe3O4 was encased in nanospheres. Thermodynamic test showed that Fe3O4 could serve as a heating source under AMF and lead the nanospheres to reach their steady temperature (45 °C). The release results in vitro indicated that nanospheres had an obvious effect of sustained release of GEM. The result of cytotoxicity assay showed that the toxicity of this material was classified as grade 1, which belongs to no cytotoxicity. The antitumor efficacy of the GEM/Fe3O4 albumin nanospheres combined with magnetic fluid hyperthermia on non-small lung cancer cell line GlC-82 was examined by MTT assay and flow cytometry assay. Compared with nanospheres entrapping GEM group, nanospheres entrapping Fe3O4 combined with MFH group, and GEM/Fe3O4 albumin nanospheres without MFH group, the GEM/Fe3O4 albumin nanospheres exhibited enhanced antitumor efficacy. Thus, the GEM/Fe3O4 albumin nanospheres have promising applications in cancer treatment.

  20. Nanovesicles from Malassezia sympodialis and host exosomes induce cytokine responses--novel mechanisms for host-microbe interactions in atopic eczema.

    Directory of Open Access Journals (Sweden)

    Ulf Gehrmann

    Full Text Available BACKGROUND: Intercellular communication can occur via the release of membrane vesicles. Exosomes are nanovesicles released from the endosomal compartment of cells. Depending on their cell of origin and their cargo they can exert different immunoregulatory functions. Recently, fungi were found to produce extracellular vesicles that can influence host-microbe interactions. The yeast Malassezia sympodialis which belongs to our normal cutaneous microbial flora elicits specific IgE- and T-cell reactivity in approximately 50% of adult patients with atopic eczema (AE. Whether exosomes or other vesicles contribute to the inflammation has not yet been investigated. OBJECTIVE: To investigate if M. sympodialis can release nanovesicles and whether they or endogenous exosomes can activate PBMC from AE patients sensitized to M. sympodialis. METHODS: Extracellular nanovesicles isolated from M. sympodialis, co-cultures of M. sympodialis and dendritic cells, and from plasma of patients with AE and healthy controls (HC were characterised using flow cytometry, sucrose gradient centrifugation, Western blot and electron microscopy. Their ability to stimulate IL-4 and TNF-alpha responses in autologous CD14, CD34 depleted PBMC was determined using ELISPOT and ELISA, respectively. RESULTS: We show for the first time that M. sympodialis releases extracellular vesicles carrying allergen. These vesicles can induce IL-4 and TNF-α responses with a significantly higher IL-4 production in patients compared to HC. Exosomes from dendritic cell and M. sympodialis co-cultures induced IL-4 and TNF-α responses in autologous CD14, CD34 depleted PBMC of AE patients and HC while plasma exosomes induced TNF-α but not IL-4 in undepleted PBMC. CONCLUSIONS: Extracellular vesicles from M. sympodialis, dendritic cells and plasma can contribute to cytokine responses in CD14, CD34 depleted and undepleted PBMC of AE patients and HC. These novel observations have implications for

  1. Optimization, in vitro cytotoxicity and penetration capability of deformable nanovesicles of paclitaxel for dermal chemotherapy in Kaposi sarcoma.

    Science.gov (United States)

    Pathak, Kamla; Sharma, Vijay; Sharma, Meenu

    2016-11-01

    Although much research has been published on ways to overcome the low oral bioavailability of paclitaxel, exploration of novel drug delivery systems that can target paclitaxel deep in to the dermal areas in AIDS-related Kaposi sarcoma (KS) have not yet been reported. Our aim was to develop deformable nanovesicles of paclitaxel capable of being used in dermal chemotherapy, especially deep into the dermal areas of AIDS related KS. Deformable nanovesicular formulations (TS1-TS15) composed of soya lecithin and span80 were prepared by the rotary evaporation sonication method within the constraints of our Box-Behnken design. The formulations were subjected to vesicle characterization and ex vivo permeation. The optimized vesicular suspension was formulated as a gel and assessed for in vitro cytotoxicity and penetration characteristics by confocal laser scanning microscopy (CLSM). TS9 with vesicle size characteristics of 185.76 ± 2.15 nm, zeta potential of -23.2 mV, deformability index = 138.02 and cumulative drug permeation of 89.80 ± 1.84% was identified as the optimized formulation. TEM revealed spherical vesicles with firm boundaries that were stable at 4 °C. TS9 was developed as carbopol 934P gel (TG) and compared with the control gel (CG) made with the pure drug (paclitaxel). TG showed significantly higher (p < 0.05) in vitro drug permeation and flux compared to the CG. In vitro cytotoxicity study on KSY-1 cell lines revealed higher IC50 (≤17) for TS against IC50 ≤19 for TG. CLSM confirmed the penetrating potential of transfersomes via TG to the dermal layers of skin, the proposed target site. Conclusively, deformable nonovesicles of paclitaxel appear as a feasible alternative to the conventional formulations of paclitaxel in the management of AIDS-related KS.

  2. Development of innovative oil-core self-organized nanovesicles prepared with chitosan and lecithin using a 2(3) full-factorial design.

    Science.gov (United States)

    Haas, Sandra Elisa; de Andrade, Cristiane; Sansone, Pedro Ernesto da Silva; Guterres, Silvia; Dalla Costa, Teresa

    2014-11-01

    The aim of this study was to develop innovative nanosystems with isopropyl myristate as the oil core of self-assembly nanovesicles constituted of chitosan and lecithin using a 2(3) factorial design. The factors analyzed were chitosan (X1, levels 4 and 8  mg/ml), oil (X2, levels 10 and 20  mg/ml) and lecithin (X3, levels 4 and 8 mg/ml). The responses evaluated were diameter, zeta potential, pH, viscosity, and backscattering analysis. The bioavailability was evaluated after oral administration of clozapine free and nanoencapsulated in rats. The diameter ranged from 0.348 to 1.5 µm for F2 (X1, 4; X2, 10; X3, 8 mg/ml) and F7 (X1, 8; X2, 20; X3, 4  mg/ml), respectively. Laser diffractometry analysis revealed only one diameter population for all batches. Zeta potential was positive, being influenced by X1 and X2/X3 association. Viscosity values were dependent on the X1 and X2 concentrations used. A structure proposed for the nanosystem consists of chitosan forming the hydrophilic shell layer that protects the core comprised of lecithin and the hydrophobic groups of oil. The AUC0-∞ was almost 3 times higher with the clozapine nanoencapsuted in relation to free drug. It was developed a new nanosystem which is able of improving the absorption of drugs.

  3. Oxidative stress and genotoxicity of an organic and an inorganic nanomaterial to Eisenia andrei: SDS/DDAB nano-vesicles and titanium silicon oxide.

    Science.gov (United States)

    Correia, Bruno; Lourenço, Joana; Marques, Sérgio; Nogueira, Verónica; Gavina, Ana; da Graça Rasteiro, Maria; Antunes, Filipe; Mendo, Sónia; Pereira, Ruth

    2017-06-01

    In the past few years the number of studies on the toxic effects of nanomaterials (NMs) in the environment increased significantly. Nonetheless, the data is still scarce, since there is a large number of NMs and new ones are being developed each day. Soils are extremely important for life, and are easily exposed to the released NMs, thus enhanced efforts are needed to study the impacts on soil biota. The objective of the present work was to determine if different concentrations of two NMs, one inorganic (TiSiO4) and other organic (nano-vesicles of sodium sodecyl sulfate/ didodecyl dimethylammonium bromide - SDS/DDAB), are genotoxic to soil invertebrates. Additionally, it was intended to understand whether, in the event of occurring, genotoxicity was caused by the incapability of the cells to deal with the oxidative stress caused by these NMs. With that purpose, Eisenia andrei were exposed for 30 days to the artificial OECD soil contaminated with different concentrations of the NMs being tested. After the exposure, coelomocytes were extracted from earthworms and DNA damage was measured by the comet assay. The activity of antioxidant enzymes (e.g. glutathione peroxidase, glutathione reductase and glutathione-S-Transferase) and lipid peroxidation were also assessed. The results showed that both NMs were genotoxic, particularly TiSiO4 for which significant DNA damages were recorded for concentrations above 444mg of TiSiO4-NM/kg of soildw. Since no statistically significant differences were found in the tested antioxidant enzymes and in lipid peroxidation, the mechanism of genotoxicity of these NMs seemed to be unrelated with oxidative stress. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Cytotoxic effects of Gemcitabine-loaded liposomes in human anaplastic thyroid carcinoma cells

    Directory of Open Access Journals (Sweden)

    Rotiroti Domenicoantonio

    2004-09-01

    Full Text Available Abstract Background Identification of effective systemic antineoplastic drugs against anaplastic thyroid carcinomas has particularly important implications. In fact, the efficacy of the chemotherapeutic agents presently used in these tumours, is strongly limited by their low therapeutic index. Methods In this study gemcitabine was entrapped within a pegylated liposomal delivery system to improve the drug antitumoral activity, thus exploiting the possibility to reduce doses to be administered in cancer therapy. The cytotoxic effects of free or liposome-entrapped gemcitabine was evaluated against a human thyroid tumour cell line. ARO cells, derived from a thyroid anaplastic carcinoma, were exposed to different concentrations of the drug. Liposomes formulations were made up of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine/cholesterol/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-MPEG (8:3:1 molar ratio. Cell viability was assessed by both trypan bleu dye exclusion assay and fluorimetric analysis of cell DNA content. Results A cytotoxic effect of free gemcitabine was present only after 72 h incubation (ARO cell mortality increased of approximately 4 fold over control at 1 μM, 7 fold at 100 μM. When gemcitabine was encapsulated in liposomes, a significant effect was observed by using lower concentrations of the drug (increased cell mortality of 2.4 fold vs. control at 0.3 μM and earlier exposure time (24 h. Conclusion These findings show that, in vitro against human thyroid cancer cells, the gemcitabine incorporation within liposomes enhances the drug cytotoxic effect with respect to free gemcitabine, thus suggesting a more effective drug uptake inside the cells. This may allow the use of new formulations with lower dosages (side effect free for the treatment of anaplastic human thyroid tumours.

  5. Low dose gemcitabine-loaded lipid nanocapsules target monocytic myeloid-derived suppressor cells and potentiate cancer immunotherapy.

    Science.gov (United States)

    Sasso, Maria Stella; Lollo, Giovanna; Pitorre, Marion; Solito, Samantha; Pinton, Laura; Valpione, Sara; Bastiat, Guillaume; Mandruzzato, Susanna; Bronte, Vincenzo; Marigo, Ilaria; Benoit, Jean-Pierre

    2016-07-01

    Tumor-induced expansion of myeloid-derived suppressor cells (MDSCs) is known to impair the efficacy of cancer immunotherapy. Among pharmacological approaches for MDSC modulation, chemotherapy with selected drugs has a considerable interest due to the possibility of a rapid translation to the clinic. However, such approach is poorly selective and may be associated with dose-dependent toxicities. In the present study, we showed that lipid nanocapsules (LNCs) loaded with a lauroyl-modified form of gemcitabine (GemC12) efficiently target the monocytic (M-) MDSC subset. Subcutaneous administration of GemC12-loaded LNCs reduced the percentage of spleen and tumor-infiltrating M-MDSCs in lymphoma and melanoma-bearing mice, with enhanced efficacy when compared to free gemcitabine. Consistently, fluorochrome-labeled LNCs were preferentially uptaken by monocytic cells rather than by other immune cells, in both tumor-bearing mice and human blood samples from healthy donors and melanoma patients. Very low dose administration of GemC12-loaded LNCs attenuated tumor-associated immunosuppression and increased the efficacy of adoptive T cell therapy. Overall, our results show that GemC12-LNCs have monocyte-targeting properties that can be useful for immunomodulatory purposes, and unveil new possibilities for the exploitation of nanoparticulate drug formulations in cancer immunotherapy.

  6. Fabrication of biomolecule copolymer hybrid nanovesicles as energy conversion systems

    Science.gov (United States)

    Ho, Dean; Chu, Benjamin; Lee, Hyeseung; Brooks, Evan K.; Kuo, Karen; Montemagno, Carlo D.

    2005-12-01

    This work demonstrates the integration of the energy-transducing proteins bacteriorhodopsin (BR) from Halobacterium halobium and cytochrome c oxidase (COX) from Rhodobacter sphaeroides into block copolymeric vesicles towards the demonstration of coupled protein functionality. An ABA triblock copolymer-based biomimetic membrane possessing UV-curable acrylate endgroups was synthesized to serve as a robust matrix for protein reconstitution. BR-functionalized polymers were shown to generate light-driven transmembrane pH gradients while pH gradient-induced electron release was observed from COX-functionalized polymers. Cooperative behaviour observed from composite membrane functionalized by both proteins revealed the generation of microamp-range currents with no applied voltage. As such, it has been shown that the fruition of technologies based upon bio-functionalizing abiotic materials may contribute to the realization of high power density devices inspired by nature.

  7. Designed biodegradable hydrogel structures prepared by stereolithography using poly(ethylene glycol)/poly(D,L-lactide)-based resins

    NARCIS (Netherlands)

    Seck, Tetsu M.; Melchels, Ferry P. W.; Feijen, Jan; Grijpma, Dirk W.

    2010-01-01

    Designed three-dimensional biodegradable poly(ethylene glycol)/poly(D,L-lactide) hydrogel structures were prepared for the first time by stereolithography at high resolutions. A photo-polymerisable aqueous resin comprising PDLLA-PEG-PDLLA-based macromer, visible light photo-initiator, dye and inhibi

  8. Preparation and Characterization of Polymeric Micelles from Poly(D,L-lactide) and Methoxypolyethylene Glycol Block Copolymers as Potential Drug Carriers

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Amphiphilic diblock copolymers composed of methoxy polyethylene glycol (MePEG) and poly(D,L-lactide) (PDLLA) were prepared for the preparation of polymeric micelles. The use of MePEG-PDLLA as drug carriers has been reported in the open literature, but there are only few data on the application of a series of MePEG-PDLLA copolymers with different lengths in the medical field. The shape of the polymeric micelles is also important in drug delivery. Studies on in vitro drug release profiles require a good sink condition. The critical micelle concentration of a series of MePEG-PDLLA has a significant role in drug release. To estimate their feasibility as a drug carrier, polymeric micelles made of MePEG-PDLLA block copolymer were prepared by the oil in water (O/W) emulsion method. From dynamic light scattering (DLS) measurements,the size of the micelle formed was less than 200 nm. The critical micelle concentration of polymeric micelles with various compositions was determined using pyrene as a fluorescence probe. The critical micelle concentration decreased with increasing number of hydrophobic segments. MePEG-PDLLA micelles have a considerably low critical micelle concentration (0.4-0.5 μg/mL), which is apparently an advantage in utilizing these micelles as drug carriers. The morphology of the polymeric micelles was observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The micelles were found to be nearly spherical. The yield of the polymeric micelles obtained from the ONV method is as high as 85%.

  9. Nanovesicle-Carbon Nanotube Hybrid Structures Mimicking Mammalian Pain Sensory System

    Science.gov (United States)

    Cho, Youngtac; Jin, Hye Jun; An, Jeong Mi; Park, Juhun; Moon, Seok Jun; Hong, Seunghun

    2015-03-01

    We developed a ``chemical-pain sensor'' based on a single-walled carbon nanotube-based field effect transistor (SWNT-FET) functionalized with rat pain sensory receptor, rat transient receptor potential vanilloid 1 (rTRPV1) mimicking a mammalian pain sensory system. The sensor can selectively detect chemical pain stimuli such as capsaicin and resiniferatoxin with a sensitivity of a 1 pM detection limit. Since this sensor allows one to quantitatively measure the concentration of chemical pain stimuli just like animal sensory systems, it can be used for various practical applications such as food screening. In addition, TRP families including rTRPV1 protein used for the sensor are now suggested as potential drug targets related to nerve and circulation disorders. Thus, the capability of measuring TRP responses using our sensor platform should open up other applications such as drug screening and basic research related with nerve and circulation systems.

  10. Natural extracellular nanovesicles and photodynamic molecules: is there a future for drug delivery?

    Science.gov (United States)

    Kusuzaki, Katsuyuki; Matsubara, Takao; Murata, Hiroaki; Logozzi, Mariantonia; Iessi, Elisabetta; Di Raimo, Rossella; Carta, Fabrizio; Supuran, Claudiu T; Fais, Stefano

    2017-12-01

    Photodynamic molecules represent an alternative approach for cancer therapy for their property (i) to be photo-reactive; (ii) to be not-toxic for target cells in absence of light; (iii) to accumulate specifically into tumour tissues; (iv) to be activable by a light beam only at the tumour site and (v) to exert cytotoxic activity against tumour cells. However, to date their clinical use is limited by the side effects elicited by systemic administration. Extracellular vesicles are endogenous nanosized-carriers that have been recently introduced as a natural delivery system for therapeutic molecules. We have recently shown the ability of human exosomes to deliver photodynamic molecules. Therefore, this review focussed on extracellular vesicles as a novel strategy for the delivery of photodynamic molecules at cancer sites. This completely new approach may enhance the delivery and decrease the toxicity of photodynamic molecules, therefore, represent the future for photodynamic therapy for cancer treatment.

  11. Size-dependent, stochastic nature of lipid exchange between nano-vesicles and model membranes

    Science.gov (United States)

    Tabaei, Seyed R.; Gillissen, Jurriaan J. J.; Vafaei, Setareh; Groves, Jay T.; Cho, Nam-Joon

    2016-07-01

    The interaction of nanoscale lipid vesicles with cell membranes is of fundamental importance for the design and development of vesicular drug delivery systems. Here, we introduce a novel approach to study vesicle-membrane interactions whereby we are able to probe the influence of nanoscale membrane properties on the dynamic adsorption, exchange, and detachment of vesicles. Using total internal reflection fluorescence (TIRF) microscopy, we monitor these processes in real-time upon the electrostatically tuned attachment of individual, sub-100 nm vesicles to a supported lipid bilayer. The observed exponential vesicle detachment rate depends strongly on the vesicle size, but not on the vesicle charge, which suggests that lipid exchange occurs during a single stochastic event, which is consistent with membrane stalk formation. The fluorescence microscopy assay developed in this work may enable measuring of the probability of stalk formation in a controlled manner, which is of fundamental importance in membrane biology, offering a new tool to understand nanoscale phenomena in the context of biological sciences.The interaction of nanoscale lipid vesicles with cell membranes is of fundamental importance for the design and development of vesicular drug delivery systems. Here, we introduce a novel approach to study vesicle-membrane interactions whereby we are able to probe the influence of nanoscale membrane properties on the dynamic adsorption, exchange, and detachment of vesicles. Using total internal reflection fluorescence (TIRF) microscopy, we monitor these processes in real-time upon the electrostatically tuned attachment of individual, sub-100 nm vesicles to a supported lipid bilayer. The observed exponential vesicle detachment rate depends strongly on the vesicle size, but not on the vesicle charge, which suggests that lipid exchange occurs during a single stochastic event, which is consistent with membrane stalk formation. The fluorescence microscopy assay developed in this work may enable measuring of the probability of stalk formation in a controlled manner, which is of fundamental importance in membrane biology, offering a new tool to understand nanoscale phenomena in the context of biological sciences. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr03817d

  12. PDE5 Inhibitors-Loaded Nanovesicles: Physico-Chemical Properties and In Vitro Antiproliferative Activity

    Directory of Open Access Journals (Sweden)

    Roberta F. De Rose

    2016-05-01

    Full Text Available Novel therapeutic approaches are required for the less differentiated thyroid cancers which are non-responsive to the current treatment. In this study we tested an innovative formulation of nanoliposomes containing sildenafil citrate or tadalafil, phosphodiesterase-5 inhibitors, on two human thyroid cancer cell lines (TPC-1 and BCPAP. Nanoliposomes were prepared by the thin layer evaporation and extrusion methods, solubilizing the hydrophilic compound sildenafil citrate in the aqueous phase during the hydration step and dissolving the lipophilic tadalafil in the organic phase. Nanoliposomes, made up of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine monohydrate (DPPC, cholesterol, and N-(carbonyl-methoxypolyethylene glycol-2000-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-mPEG2000 (6:3:1 molar ratio, were characterized by a mean diameter of ~100 nm, a very low polydispersity index (~0.1 and a negative surface charge. The drugs did not influence the physico-chemical properties of the systems and were efficiently retained in the colloidal structure. By using cell count and MTT assay, we found a significant reduction of the viability in both cell lines following 24 h treatment with both nanoliposomal-encapsulated drugs, notably greater than the effect of the free drugs. Our findings demonstrate that nanoliposomes increase the antiproliferative activity of phosphodiesterase-5 inhibitors, providing a useful novel formulation for the treatment of thyroid carcinoma.

  13. Exosomes: a role for naturally occurring nanovesicles in cancer growth, diagnosis and treatment.

    Science.gov (United States)

    Srivastava, Akhil; Filant, Justyna; Moxley, Katherine M; Sood, Anil; McMeekin, Scott; Ramesh, Rajagopal

    2015-01-01

    Exosomes are 30-100 nm bodies secreted from almost all types of cells into the extracellular spaces. They enclose in their lumen active genetic information in the form of messenger RNA (mRNA), micro RNA (miRNA), DNA and active peptides that are representative of the parental cell and can be isolated from different body fluids. Exosomes can participate in inter-cellular communication by trafficking molecules to their target cells. Because they can stably carry cargo including miRNA, mRNA, and proteins and can pass through stringent biological barriers (e.g., blood brain barrier) without eliciting an immune response, they are considered as an ideal acellular vehicle for drug delivery. In this review, we describe the structure and biogenesis of exosomes and new directions related to their role in diagnosis and treatment of diseases, especially for cancer. We also discuss potential challenges associated with exosomes that should be addressed before exosome-based therapy can be applied to clinical settings.

  14. Study of natural nanovesicles carrying olfactory receptors for the development of biosensing platforms

    OpenAIRE

    Sanmartí Espinal, Marta

    2015-01-01

    [eng] Natural vesicles produced from genetically engineered cells with tailored membrane receptor composition are promising building blocks for sensing biodevices. This is particularly true for the case of G-protein coupled receptors (GPCRs) present in many sensing processes in cells, whose functionality crucially depends on their lipid environment. Membrane receptors are involved in a variety of biochemical pathways and therefore constitute important targets for therapy and development of ne...

  15. Chitosan/lecithin liposomal nanovesicles as an oral insulin delivery system.

    Science.gov (United States)

    Al-Remawi, Mayyas; Elsayed, Amani; Maghrabi, Ibrahim; Hamaidi, Mohammad; Jaber, Nisrein

    2017-05-01

    In the present work, insulin-chitosan polyelectrolyte complexes associated to lecithin liposomes were investigated as a new carrier for oral delivery of insulin. The preparation was characterized in terms of particle size, zeta potential and encapsulation efficiency. Surface tension measurements revealed that insulin-chitosan polyelectrolyte complexes have some degree of hydrophobicity and should be added to lecithin liposomal dispersion and not the vice versa to prevent their adsorption on the surface. Stability of insulin was enhanced when it was associated to liposomes. Significant reduction of blood glucose levels was noticed after oral administration of liposomal preparation to streptozotocin diabetic rats compared to control. The hypoglycemic activity was more prolonged compared to subcutaneously administered insulin.

  16. Nanovesicle-targeted Kv1.3 knockdown in memory T cells suppresses CD40L expression and memory phenotype.

    Science.gov (United States)

    Chimote, Ameet A; Hajdu, Peter; Kottyan, Leah C; Harley, John B; Yun, Yeoheung; Conforti, Laura

    2016-05-01

    Ca(2+) signaling controls activation and effector functions of T lymphocytes. Ca(2+) levels also regulate NFAT activation and CD40 ligand (CD40L) expression in T cells. CD40L in activated memory T cells binds to its cognate receptor, CD40, on other cell types resulting in the production of antibodies and pro-inflammatory mediators. The CD40L/CD40 interaction is implicated in the pathogenesis of autoimmune disorders and CD40L is widely recognized as a therapeutic target. Ca(2+) signaling in T cells is regulated by Kv1.3 channels. We have developed lipid nanoparticles that deliver Kv1.3 siRNAs (Kv1.3-NPs) selectively to CD45RO(+) memory T cells and reduce the activation-induced Ca(2+) influx. Herein we report that Kv1.3-NPs reduced NFAT activation and CD40L expression exclusively in CD45RO(+) T cells. Furthermore, Kv1.3-NPs suppressed cytokine release and induced a phenotype switch of T cells from predominantly memory to naïve. These findings indicate that Kv1.3-NPs operate as targeted immune suppressive agents with promising therapeutic potentials.

  17. Transdermal glimepiride delivery system based on optimized ethosomal nano-vesicles: Preparation, characterization, in vitro, ex vivo and clinical evaluation.

    Science.gov (United States)

    Ahmed, Tarek A; El-Say, Khalid M; Aljaeid, Bader M; Fahmy, Usama A; Abd-Allah, Fathy I

    2016-03-16

    This work aimed to develop an optimized ethosomal formulation of glimepiride then loading into transdermal films to offer lower drug side effect, extended release behavior and avoid first pass effect. Four formulation factors were optimized for their effects on vesicle size (Y1), entrapment efficiency (Y2) and vesicle flexibility (Y3). Optimum desirability was identified and, an optimized formulation was prepared, characterized and loaded into transdermal films. Ex-vivo permeation study for the prepared films was conducted and, the permeation parameters and drug permeation mechanism were identified. Penetration through rat skin was studied using confocal laser microscope. In-vivo study was performed following transdermal application on human volunteers. The percent of alcohol was significantly affecting all the studied responses while the other factors and their interaction effects were varied on their effects on each response. The optimized ethosomal formulation showed observed values for Y1, Y2 and Y3 of 61 nm, 97.12% and 54.03, respectively. Ex-vivo permeation of films loaded with optimized ethosomal formulation was superior to that of the corresponding pure drug transdermal films and this finding was also confirmed after confocal laser microscope study. Permeation of glimepiride from the prepared films was in favor of Higushi-diffusion model and exhibited non-Fickian or anomalous release mechanism. In-vivo study revealed extended drug release behavior and lower maximum drug plasma level from transdermal films loaded with drug ethosomal formulation. So, the ethosomal formulation could be considered a suitable drug delivery system especially when loaded into transdermal vehicle with possible reduction in side effects and controlling the drug release.

  18. Toward targeted therapy in chemotherapy-resistant pancreatic cancer with a smart triptolide nanomedicine.

    Science.gov (United States)

    Wang, Cheng; Liu, Biao; Xu, Xuelian; Zhuang, Bo; Li, Hongxia; Yin, Jiaqi; Cong, Mengyi; Xu, Wei; Lu, Aiping

    2016-02-16

    Chemoresistance is the major impediment for treating pancreatic cancer. Herb-derived compound triptolide (TP) can inhibit proliferation of chemo-resistant pancreatic cancer (CPC) cell lines through multiple mechanisms, which exhibited superior anticancer efficacy compared with gemcitabine. However, toxicity due to non-specific exposure to healthy tissues hindered its clinical translation. Herein we successfully achieved targeting CPC cells and avoiding exposure to healthy tissues for TP by nucleolin-specific aptamer (AS1411) mediated polymeric nanocarrier. We conjugated AS1411 aptamer to carboxy terminated poly(ethylene glycol)-block-poly(d, l-lactide) (HOOC-PEG-PDLLA), then prepared AS1411-PEG-PDLLA micelle loading TP (AS-PPT) through solid dispersion technique. AS-PPT showed more antitumor activity than TP and equivalent specific binding ability with gemcitabine-resistant human pancreatic cancer cell (MIA PaCa-2) to AS1411 aptamer in vitro. Furthermore, we studied the distribution of AS-PPT (Cy3-labed TP) at tissue and cellular levels using biophotonic imaging technology. The results showed AS1411 facilitated TP selectively accumulating in tumor tissues and targeting CPC cells. The lifetime of the MIA PaCa-2 cell-bearing mice administrated with AS-PPT was efficiently prolonged than that of the mice subjected to the clinical anticancer drug Gemzar® in vivo. Such work provides a new strategy for overcoming the drug resistance of pancreatic cancer.

  19. Construction of Epidermal Growth Factor Receptor Peptide Magnetic Nanovesicles with Lipid Bilayers for Enhanced Capture of Liver Cancer Circulating Tumor Cells.

    Science.gov (United States)

    Ding, Jian; Wang, Kai; Tang, Wen-Jie; Li, Dan; Wei, You-Zhen; Lu, Ying; Li, Zong-Hai; Liang, Xiao-Fei

    2016-09-20

    Highly effective targeted tumor recognition via vectors is crucial for cancer detection. In contrast to antibodies and proteins, peptides are direct targeting ligands with a low molecular weight. In the present study, a peptide magnetic nanovector platform containing a lipid bilayer was designed using a peptide amphiphile (PA) as a skeleton material in a controlled manner without surface modification. Fluorescein isothiocyanate-labeled epidermal growth factor receptor (EGFR) peptide nanoparticles (NPs) could specifically bind to EGFR-positive liver tumor cells. EGFR peptide magnetic vesicles (EPMVs) could efficiently recognize and separate hepatoma carcinoma cells from cell solutions and treated blood samples (ratio of magnetic EPMVs versus anti-EpCAM NPs: 3.5 ± 0.29). Analysis of the circulating tumor cell (CTC) count in blood samples from 32 patients with liver cancer showed that EPMVs could be effectively applied for CTC capture. Thus, this nanoscale, targeted cargo-packaging technology may be useful for designing cancer diagnostic systems.

  20. Development of tumor-specific caffeine-potentiated chemotherapy using a novel drug delivery system with Span 80 nano-vesicles

    OpenAIRE

    NAKATA, HIROSHI; Miyazaki, Tatsuhiko; Iwasaki, Tomoyuki; Nakamura, Atsushi; Kidani, Teruki; Sakayama, Kenshi; Masumoto, Junya; MIURA, HIROMASA

    2015-01-01

    In recent years, chemotherapy with caffeine has manifested potently high efficacy against osteosarcoma, although adverse effects have been observed. Recently, we developed a novel drug delivery system (DDS) with nonionic vesicles prepared from Span 80 which have promising physicochemical properties as an attractive possible alternative to commonly used liposomes. Herein, we demonstrated that tumor-specific caffeine-potentiated chemotherapy for murine osteosarcoma administered by a novel DDS w...

  1. Designed biodegradable hydrogel structures prepared by stereolithography using poly(ethylene glycol)/poly(D,L-lactide)-based resins.

    Science.gov (United States)

    Seck, Tetsu M; Melchels, Ferry P W; Feijen, Jan; Grijpma, Dirk W

    2010-11-20

    Designed three-dimensional biodegradable poly(ethylene glycol)/poly(D,L-lactide) hydrogel structures were prepared for the first time by stereolithography at high resolutions. A photo-polymerisable aqueous resin comprising PDLLA-PEG-PDLLA-based macromer, visible light photo-initiator, dye and inhibitor in DMSO/water was used to build the structures. Porous and non-porous hydrogels with well-defined architectures and good mechanical properties were prepared. Porous hydrogel structures with a gyroid pore network architecture showed narrow pore size distributions, excellent pore interconnectivity and good mechanical properties. The structures showed good cell seeding characteristics, and human mesenchymal stem cells adhered and proliferated well on these materials.

  2. Platelet-Rich Plasma-Loaded Poly(d,l-lactide-Poly(ethylene glycol-Poly(d,l-lactide Hydrogel Dressing Promotes Full-Thickness Skin Wound Healing in a Rodent Model

    Directory of Open Access Journals (Sweden)

    Manle Qiu

    2016-06-01

    Full Text Available Traditional therapeutic methods for skin wounds have many disadvantages, and new wound dressings that can facilitate the healing process are thus urgently needed. Platelet-rich plasma (PRP contains multiple growth factors (GFs and shows a significant capacity to heal soft tissue wounds. However, these GFs have a short half-life and deactivate rapidly; we therefore need a sustained delivery system to overcome this shortcoming. In this study, poly(d,l-lactide-poly(ethylene glycol-poly(d,l-lactide (PDLLA-PEG-PDLLA: PLEL hydrogel was successfully created as delivery vehicle for PRP GFs and was evaluated systematically. PLEL hydrogel was injectable at room temperature and exhibited a smart thermosensitive in situ gel-formation behavior at body temperature. In vitro cell culture showed PRP-loaded PLEL hydrogel (PRP/PLEL had little cytotoxicity, and promoted EaHy926 proliferation, migration and tube formation; the factor release assay additionally indicated that PLEL realized the controlled release of PRP GFs for as long as 14 days. When employed to treat rodents’ full-thickness skin defects, PRP/PLEL showed a significantly better ability to raise the number of both newly formed and mature blood vessels compared to the control, PLEL and PRP groups. Furthermore, the PRP/PLEL-treated group displayed faster wound closure, better reepithelialization and collagen formation. Taken together, PRP/PLEL provides a promising strategy for promoting angiogenesis and skin wound healing, which extends the potential of this dressing for clinical application.

  3. FG020326-1oaded nanoparticle with PEG and PDLLA improved pharma-codynamics of reversing multidrug resistance in vitro and in vivo

    Institute of Scientific and Technical Information of China (English)

    Wen-jing DENG; Xiao-qiang YANG; Yong-ju LIANG; Li-ming CHEN; Yan-yan YAN; Xin-tao SHUAI; Li-wu FU

    2007-01-01

    Aim: FG020326, a novel imidazole derivative, is a potent multidrug-resistance (MDR) modulator in vitro and in vivo. However, FG020326 is insoluble. PEDLLA-FG020326 is a FG020326-loaded nanoparticle formed with diblock eopolymers of poly (ethylene glycol)-bloek-poly (D,L-lactic acid) (PEG:PDLLA, PEDLLA) that can solubilize FG020326. This work was intended to evaluate the pharmacody-namics of PEDLLA-FG020326 on reversing MDR in vitro and in vivo.Methods: Cytotoxicity was determined by tetrazolium assay. The intraceUular accumulation and efflux of doxorubicin (Dox) were detected by fluorescence spectrophotometry.The function of P-glycoprotein was examined by Rhodamine 123 (Rh 123) accumu-lation detected by flow cytometry. The KBv200 cell xenograft model was estab-lished to investigate the effect of PEDLLA-FG020326 on reversing MDR in vivo.Results: PEDLLA-FG020326 and FG020326 exhibited 56.4- and 35.9-fold activity in reversing KBv200 cells to vincristine (VCR) resistance, respectively and 14.98-and 7.64-fold to Dox resistance, respectively. PEDLLA-FG020326 was much stron-ger than FG020326, resulting in the increase of Dox and Rh123 accumulation and the decrease of intracellular Dox extrusion in KBv200 cells. Importantly, PEDLLA-FG020326 exhibited more powerful activity than FG020326 in enhancing the effect of VCR against KBv200 cell xenografts in nude mice, but did not appear more toxic.Conclusion: The pharmacodynamics of FG020326 was improved by incorporating it into a micellar nanoparticle formed with PEG-block-PDLLA copolymers.

  4. Synthesis, characterization and third-order nonlinear optical properties of polydiacetylene nanostructures, silver nanoparticles and polydiacetylene–silver nanocomposites

    Indian Academy of Sciences (India)

    B BHUSHAN; S S TALWAR; T KUNDU; B P SINGH

    2016-10-01

    We have synthesized, characterized and studied the third-order nonlinear optical properties of two different nanostructures of polydiacetylene (PDA), PDA nanocrystals and PDA nanovesicles, along with silver nanoparticles-decorated PDA nanovesicles. The second molecular hyperpolarizability $\\gamma (−\\omega; \\omega,−\\omega,\\omega$) of the samples has been investigated by antiresonant ring interferometric nonlinear spectroscopic (ARINS) technique using femtosecond mode-locked Ti:sapphire laser in the spectral range of 720–820 nm. The observed spectral dispersion of $\\gamma$ has been explained in the framework of three-essential states model and a correlation between the electronic structure and optical nonlinearity of the samples has been established. The energy of two-photon state, transition dipole moments and linewidth of the transitions have been estimated. We have observed that the nonlinear optical properties of PDA nanocrystals and nanovesicles are different because of the influence of chain coupling effects facilitated by the chain packing geometry of the monomers. On the other hand, our investigation reveals that the spectral dispersion characteristic of $\\gamma$ for silver nanoparticles-coated PDA nanovesicles is qualitatively similar to that observed for the uncoated PDA nanovesicles but bears no resemblance to that observed in silver nanoparticles. The presence of silver nanoparticles increases the $\\gamma$ values of the coated nanovesicles slightly as compared to that of the uncoated nanovesicles, suggesting a definite but weak coupling between the free electrons of the metal nanoparticles and $\\pi$ electrons of the polymer in the composite system. Our comparative studies show that the arrangement of polymer chains in polydiacetylene nanocrystals is more favourable for higher nonlinearity.

  5. Organic Nanovesicular Cargoes for Sustained Drug Delivery: Synthesis, Vesicle Formation, Controlling “Pearling” States, and Terfenadine Loading/Release Studies

    Directory of Open Access Journals (Sweden)

    Ajay Kumar Botcha

    2014-01-01

    Full Text Available “Sustained drug delivery systems” which are designed to accomplish long-lasting therapeutic effect are one of the challenging topics in the area of nanomedicine. We developed an innovative strategy to prepare nontoxic and polymer stabilized organic nanovesicles (diameter: 200 nm from a novel bolaamphiphile, where two hydrogen bonding acetyl cytosine molecules connected to 4,4′′-positions of the 2,6-bispyrazolylpyridine through two flexible octyne chains. The nanovesicles behave like biological membrane by spontaneously self-assembling into “pearl-like” chains and subsequently forming long nanotubes (diameter: 150 nm, which further develop into various types of network-junctions through self-organization. For drug loading and delivery applications, the nanovesicles were externally protected with biocompatible poly(ethyleneglycol-2000 to prevent them from fusion and ensuing tube formation. Nontoxic nature of the nanovesicles was demonstrated by zebrafish teratogenicity assay. Biocompatible nanovesicles were loaded with “terfenadine” drug and successfully utilized to transport and release drug in sustained manner (up to 72 h in zebrafish larvae, which is recognized as an emerging in vivo model system.

  6. Stability of membranous nanostructures: a possible key mechanism in cancer progression

    Directory of Open Access Journals (Sweden)

    Kralj-Iglic V

    2012-07-01

    Full Text Available Veronika Kralj-IglicBiomedical Research Group, Faculty of Health Sciences, University of Ljubljana, Zdravstvena 5, Ljubljana, SloveniaAbstract: Membranous nanostructures, such as nanovesicles and nanotubules, are an important pool of biological membranes. Recent results indicate that they constitute cell-cell communication systems and that cancer development is influenced by these systems. Nanovesicles that are pinched off from cancer cells can move within the circulation and interact with distant cells. It has been suggested and indicated by experimental evidence that nanovesicles can induce metastases from the primary tumor in this way. Therefore, it is of importance to understand better the mechanisms of membrane budding and vesiculation. Here, a theoretical description is presented concerning consistently related lateral membrane composition, orientational ordering of membrane constituents, and a stable shape of nanovesicles and nanotubules. It is shown that the character of stable nanostructures reflects the composition of the membrane and the intrinsic shape of its constituents. An extension of the fluid mosaic model of biological membranes is suggested by taking into account curvature-mediated orientational ordering of the membrane constituents on strongly anisotropically curved regions. Based on experimental data for artificial membranes, a possible antimetastatic effect of plasma constituents via mediation of attractive interaction between membranous structures is suggested. This mediated attractive interaction hypothetically suppresses nanovesiculation by causing adhesion of buds to the mother membrane and preventing them from being pinched off from the membrane.Keywords: nanovesicles, nanotubules, nanotubes, microvesicles, exosomes, metastasis

  7. Polysaccharide nano-vesicular multidrug carriers for synergistic killing of cancer cells

    Science.gov (United States)

    Pramod, P. S.; Shah, Ruchira; Chaphekar, Sonali; Balasubramanian, Nagaraj; Jayakannan, Manickam

    2014-09-01

    Multi-drug delivery based on polymer nano-scaffolds is an essential protocol to be developed for better administration of anticancer drugs to enhance their therapeutic efficacies against cancer cells. Here, we report dual delivery polysaccharide nano-vesicles that are capable of loading and delivering both water soluble and water insoluble drugs together in a single polymer scaffold. The selective rupture of the nano-vesicular assembly under intracellular enzyme conditions allowed the simultaneous delivery of a hydrophobic drug camptothecin (CPT) and hydrophilic drug doxorubicin (DOX) supporting their synergistic killing of breast and colon cancer cells. The polysaccharide nano-vesicles have allowed us to address a few important questions regarding the need for multiple drug administration in cancer cells including (a) the role of simultaneous drug release, (b) antagonistic versus synergistic effects of drug combinations and (c) how these are affected by the ratio of drugs. Further, evaluation of the role of caveolae in endocytosis of these polymer scaffolds was also made. The vesicular scaffolds were found to preserve and deliver DOX resulting in 50-60% better killing of cancer cells than the free drug. Additionally, dual loaded nano-vesicles when compared to drug cocktails with individual drugs in separate nano-vesicles (at comparable molar ratios) suggest the relative drug concentration following release and mode of delivery to be both important in cancer cell killing. Results from these experiments have revealed newly developed polysaccharide nano-vesicles loaded with DOX and CPT drugs as potential candidates for improved breast cancer cell killing. Thus, these custom-designed polysaccharide nano-vesicles provide a new perspective on multi-anticancer drug delivery systems and their efficacy.Multi-drug delivery based on polymer nano-scaffolds is an essential protocol to be developed for better administration of anticancer drugs to enhance their therapeutic

  8. Control of protein adsorption on functionalized electrospun fibers.

    Science.gov (United States)

    Grafahrend, Dirk; Calvet, Julia Lleixa; Klinkhammer, Kristina; Salber, Jochen; Dalton, Paul D; Möller, Martin; Klee, Doris

    2008-10-15

    Electrospun fibers that are protein resistant and functionalized with bioactive signals were produced by solution electrospinning amphiphilic block copolymers. Poly (ethylene glycol)-block-poly(D,L-lactide) (PEG-b-PDLLA) was synthesized in two steps, with a PEG segment of 10 kDa, while the PDLLA block ranged from 20 to 60 kDa. Depending on the PEG and PDLLA segment ratio, as well as solvent selection, the hydrophilicity and protein adsorption could be altered on the electrospun mesh. Furthermore, an alpha-acetal PEG-b-PDLLA was synthesized that allowed the conjugation of active molecules, resulting in surface functionalization of the electrospun fiber. Electrospun material with varying morphologies and diameter were electrospun from 10, 20, and 30 wt.% solutions. Sessile drop measurements showed a reduction in the contact angle from 120 degrees for pure poly(D,L-lactide) with increasing PEG/PDLLA ratio. All electrospun block PEG-b-PDLLA fibers had hydrophilic properties, with contact angles below 45 degrees . The fibers were collected onto six-arm star-poly(ethylene glycol) (star-PEG) coated silicon wafers and incubated with fluorescently labeled proteins. All PEG-b-PDLLA fibers showed no detectable adsorption of bovine serum albumin (BSA) independent of their composition while a dependence between hydrophobic block length was observed for streptavidin adsorption. Fibers of block copolymers with PDLLA blocks smaller than 39 kDa showed no adsorption of BSA or streptavidin, indicating good non-fouling properties. Fibers were surface functionalized with N(epsilon)-(+)-biotinyl-L-lysine (biocytin) or RGD peptide by attaching the molecule to the PEG block during synthesis. Protein adsorption measurements, and the controlled interaction of biocytin with fluorescently labeled streptavidin, showed that the electrospun fibers were both resistant to protein adsorption and are functionalized. Fibroblast adhesion was contrasting between the unfunctionalized and RGD

  9. Targeting Ovarian Cancer with Porphysome Nanotechnology

    Science.gov (United States)

    2016-10-01

    included. 24-hours post -injection the mice receiving Porphysomes are sacrificed and tissue biodistribution performed using ex Vivo tissue homogenate...human OC. Body: Preparation of Porphysome Nanovesicles with Reverse-Phase Evaporation (Figure 2). The production of Porphysomes had been previously...reported, following a multistep thin film hydration (TFH) procedure commonly used for conventional liposomal preparations [1]. Briefly, dried lipidic

  10. Modulation of tissue tropism and biological activity of exosomes and other extracellular vesicles : New nanotools for cancer treatment

    NARCIS (Netherlands)

    Kooijmans, Sander A A; Schiffelers, Raymond M.; Zarovni, Natasa; Vago, Riccardo

    2016-01-01

    Exosomes are naturally secreted nanovesicles that have recently aroused a great interest in the scientific and clinical community for their roles in intercellular communication in almost all physiological and pathological processes. These 30–100 nm sized vesicles are released from the cells into the

  11. Increased PSA expression on prostate cancer exosomes in in vitro condition and in cancer patients.

    Science.gov (United States)

    Logozzi, Mariantonia; Angelini, Daniela F; Iessi, Elisabetta; Mizzoni, Davide; Di Raimo, Rossella; Federici, Cristina; Lugini, Luana; Borsellino, Giovanna; Gentilucci, Alessandro; Pierella, Federico; Marzio, Vittorio; Sciarra, Alessandro; Battistini, Luca; Fais, Stefano

    2017-09-10

    Prostate specific antigen (PSA) test is the most common, clinically validated test for the diagnosis of prostate cancer (PCa). While neoplastic lesions of the prostate may cause aberrant levels of PSA in the blood, the quantitation of free or complexed PSA poorly discriminates cancer patients from those developing benign lesions, often leading to invasive and unnecessary surgical procedures. Microenvironmental acidity increases exosome release by cancer cells. In this study we evaluated whether acidity, a critical phenotype of malignancy, could influence exosome release and increase the PSA expression in nanovesicles released by PCa cells. To this aim, we exploited Nanoparticle Tracking Analysis (NTA), an immunocapture-based ELISA, and nanoscale flow-cytometry. The results show that microenvironmental acidity induces an increased release of nanovesicles expressing both PSA and the exosome marker CD81. In order to verify whether the changes induced by the local selective pressure of extracellular acidity may correspond to a clinical pathway we used the same approach to evaluate the levels of PSA-expressing exosomes in the plasma of PCa patients and controls, including subjects with benign prostatic hypertrophy (BPH). The results show that only PCa patients have high levels of nanovesicles expressing both CD81 and PSA. This study shows that tumor acidity exerts a selective pressure leading to the release of extracellular vesicles that express both PSA and exosome markers. A comparable scenario was shown in the plasma of prostate cancer patients as compared to both BPH and healthy controls. These results suggest that microenvironmental acidity may represent a key factor which determines qualitatively and quantitatively the release of extracellular vesicles by malignant tumors, including prostate cancer. This condition leads to the spill-over of nanovesicles into the peripheral blood of prostate cancer patients, where the levels of tumor biomarkers expressed by

  12. Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin.

    Science.gov (United States)

    Ho, Tien Yu Jessica; Chan, Chia-Chung; Chan, KinGho; Wang, Yu Chieh; Lin, Jing-Tang; Chang, Cheng-Ming; Chen, Chien-Sheng

    2013-11-15

    We developed a sensitive, simple, inexpensive and rapid bead-based immunoassay platform, composed of liposomal nanovesicle amplification system, Gentamycin sulfate beads and 96-well filtration plates. In the beginning of the assay, Gentamycin sulfate beads, Gentamycin sulfate and Gentamycin specific antibody were incubated in a bottom-sealed 96-well filtration plate. After incubation, washing was done by running washing buffer through the unsealed filtration plate with only gravity and the antibody-Gentamycin bead complexes were retained in the plate. Fluorescent dye-loaded protein G-liposomal nanovesicles were then added to specifically bind to antibodies on the retained beads. After washing unbound nanovesicles, millions of fluorescent dye molecules were released by adding a detergent solution to lyse liposomal nanovesicles. The limit of detection (LOD) of this novel detection platform in TBS and in skim milk were 52.65 ng/mL and 14.16 ng/mL, which are both sufficient for detecting the 200 ng/mL Codex maximum residual level (MRL). The dynamic ranges were both from each of their LODs to 100 μg/mL. The 50% inhibition concentrations (IC50) in TBS and skim milk were 199.66 ng/mL and 360.81 ng/mL, respectively. We also demonstrated the good specificity of this platform by comparing detection results between pure Gentamycin solution and a mixture solution of 6 different antibiotics including Gentamycin in skim milk. The entire assay with 60 samples was conducted within 2h. In sum, this novel biosensing platform not only fulfilled most benefits of magnetic bead-based assays, but also was inexpensive and convenient by replacing the magnetic separation with filtration plate separation.

  13. Therapeutic and cosmeceutical potential of ethosomes: An overview

    OpenAIRE

    Poonam Verma; Pathak, K

    2010-01-01

    The main disadvantage of transdermal drug delivery is the poor penetration of most compounds into the human skin. The main barrier of the skin is located within its uppermost layer, the stratum corneum (SC). Several approaches have been developed to weaken this skin barrier. One of the approaches for increasing the skin penetration of drugs and many cosmetic chemicals is the use of vesicular systems, such as, liposomes and ethosomes. Ethosomes are phospholipid-based elastic nanovesicles conta...

  14. Digital Detection of Exosomes by Interferometric Imaging

    OpenAIRE

    2016-01-01

    Exosomes, which are membranous nanovesicles, are actively released by cells and have been attributed to roles in cell-cell communication, cancer metastasis, and early disease diagnostics. The small size (30–100 nm) along with low refractive index contrast of exosomes makes direct characterization and phenotypical classification very difficult. In this work we present a method based on Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) that allows multiplexed phenotyping and ...

  15. Chitosan–Pluronic nanoparticles as oral delivery of anticancer gemcitabine: preparation and in vitro study

    Directory of Open Access Journals (Sweden)

    Ostad SN

    2012-04-01

    Full Text Available Hosniyeh Hosseinzadeh1, Fatemeh Atyabi1, Rassoul Dinarvand1, Seyed Naser Ostad21Nanotechnology Research Centre, 2Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, IranAbstract: Nanoparticles have proven to be an effective delivery system with few side effects for anticancer drugs. In this study, gemcitabine-loaded nanoparticles have been prepared by an ionic gelation method using chitosan and Pluronic® F-127 as a carrier. Prepared nanoparticles were characterized using dynamic light scattering, Fourier transform infrared spectroscopy (FT-IR, differential scanning calorimetry (DSC, scanning electron microscopy, and transmission electron microscopy. Different parameters such as concentration of sodium tripolyphosphate, chitosan, Pluronic, and drug on the properties of the prepared nanoparticles were evaluated. In vitro drug release was studied in phosphate-buffered saline (PBS; pH = 7.4. The cytotoxicity of the nanoparticles was assayed in the HT-29 colon cancer cell line. The mucoadhesion behavior of the nanoparticles was also studied by mucus glycoprotein assay. The prepared nanoparticles had a spherical shape with positive charge and a mean diameter ranging between 80 to 170 nm. FT-IR and DSC studies found that the drug was dispersed in its amorphous form due to its potent interaction with nanoparticle matrix. Maximum drug encapsulation efficiency was achieved at 0.4 mg/mL gemcitabine while maximum drug loading was 6% obtained from 0.6 mg/mL gemcitabine. An in vitro drug release study at 37°C in PBS (pH = 7.4 exhibited a controlled release profile for chitosan–Pluronic® F-127 nanoparticles. A cytotoxicity assay of gemcitabine-loaded nanoparticles showed an increase in the cytotoxicity of gemcitabine embedded in the nanoparticles in comparison with drug alone. The mucoadhesion study results suggest that nanoparticles could be considered as an efficient oral formulation for colon

  16. Role of extracellular vesicles in de novo mineralization: an additional novel mechanism of cardiovascular calcification.

    Science.gov (United States)

    New, Sophie E P; Aikawa, Elena

    2013-08-01

    Extracellular vesicles are membrane micro/nanovesicles secreted by many cell types into the circulation and the extracellular milieu in physiological and pathological conditions. Evidence suggests that extracellular vesicles, known as matrix vesicles, play a role in the mineralization of skeletal tissue, but emerging ultrastructural and in vitro studies have demonstrated their contribution to cardiovascular calcification as well. Cells involved in the progression of cardiovascular calcification release active vesicles capable of nucleating hydroxyapatite on their membranes. This review discusses the role of extracellular vesicles in cardiovascular calcification and elaborates on this additional mechanism of calcification as an alternative pathway to the currently accepted mechanism of biomineralization via osteogenic differentiation.

  17. Force balance and membrane shedding at the Red Blood Cell surface

    OpenAIRE

    Sens, Pierre; Gov, Nir

    2006-01-01

    During the aging of the red-blood cell, or under conditions of extreme echinocytosis, membrane is shed from the cell plasma membrane in the form of nano-vesicles. We propose that this process is the result of the self-adaptation of the membrane surface area to the elastic stress imposed by the spectrin cytoskeleton, via the local buckling of membrane under increasing cytoskeleton stiffness. This model introduces the concept of force balance as a regulatory process at the cell membrane, and qu...

  18. Exosomes released by melanoma cells prepare sentinel lymph nodes for tumor metastasis.

    Science.gov (United States)

    Hood, Joshua L; San, Roman Susana; Wickline, Samuel A

    2011-06-01

    Exosomes are naturally occurring biological nanovesicles utilized by tumors to communicate signals to local and remote cells and tissues. Melanoma exosomes can incite a proangiogenic signaling program capable of remodeling tissue matrices. In this study, we show exosome-mediated conditioning of lymph nodes and define microanatomic responses that license metastasis of melanoma cells. Homing of melanoma exosomes to sentinel lymph nodes imposes synchronized molecular signals that effect melanoma cell recruitment, extracellular matrix deposition, and vascular proliferation in the lymph nodes. Our findings highlight the pathophysiologic role and mechanisms of an exosome-mediated process of microanatomic niche preparation that facilitates lymphatic metastasis by cancer cells.

  19. Preparation of plasmonic vesicles from amphiphilic gold nanocrystals grafted with polymer brushes

    Science.gov (United States)

    Song, Jibin; Huang, Peng; Chen, Xiaoyuan

    2016-01-01

    Gold nanovesicles contain multiple nanocrystals within a polymeric coating. The strong plasmonic coupling between adjacent nanoparticles in their vesicular shell makes ultrasensitive biosensing and bioimaging possible. In our laboratory, multifunctional plasmonic vesicles are assembled from amphiphilic gold nanocrystals (such as gold nanoparticles and gold nanorods) coated with mixed hydrophilic and hydrophobic polymer brushes or amphiphilic diblock co-polymer brushes. To fulfill the different requirements of biomedical applications, different polymers that are either pH=responsive, photoactive or biodegradable can be used to form the hydrophobic brush, while the hydrophilicity is maintained by polyethylene glycol (PEG). This protocol covers the preparation, surface functionalization and self-assembly of amphiphilic gold nanocrystals grafted covalently with polymer brushes. The protocol can be completed within 2 d. The preparation of amphiphilic gold nanocrystals, coated with amphiphilic diblock polymer brushes using a ‘grafting to’ method or mixed hydrophilic and hydrophobic polymer brushes using tandem ‘grafting to’ and ‘grafting from’ methods, is described. We also provide detailed procedures for the preparation and characterization of pH-responsive plasmonic gold nanovesicles from amphiphilic gold nanocrystals using a film-rehydration method that can be completed within ~3 d. PMID:27763624

  20. Bioelectronic tongue using heterodimeric human taste receptor for the discrimination of sweeteners with human-like performance.

    Science.gov (United States)

    Song, Hyun Seok; Jin, Hye Jun; Ahn, Sae Ryun; Kim, Daesan; Lee, Sang Hun; Kim, Un-Kyung; Simons, Christopher T; Hong, Seunghun; Park, Tai Hyun

    2014-10-28

    The sense of taste helps humans to obtain information and form a picture of the world by recognizing chemicals in their environments. Over the past decade, large advances have been made in understanding the mechanisms of taste detection and mimicking its capability using artificial sensor devices. However, the detection capability of previous artificial taste sensors has been far inferior to that of animal tongues, in terms of its sensitivity and selectivity. Herein, we developed a bioelectronic tongue using heterodimeric human sweet taste receptors for the detection and discrimination of sweeteners with human-like performance, where single-walled carbon nanotube field-effect transistors were functionalized with nanovesicles containing human sweet taste receptors and used to detect the binding of sweeteners to the taste receptors. The receptors are heterodimeric G-protein-coupled receptors (GPCRs) composed of human taste receptor type 1 member 2 (hTAS1R2) and human taste receptor type 1 member 3 (hTAS1R3), which have multiple binding sites and allow a human tongue-like broad selectivity for the detection of sweeteners. This nanovesicle-based bioelectronic tongue can be a powerful tool for the detection of sweeteners as an alternative to labor-intensive and time-consuming cell-based assays and the sensory evaluation panels used in the food and beverage industry. Furthermore, this study also allows the artificial sensor to exam the functional activity of dimeric GPCRs.

  1. Real-time monitoring of geosmin and 2-methylisoborneol, representative odor compounds in water pollution using bioelectronic nose with human-like performance.

    Science.gov (United States)

    Son, Manki; Cho, Dong-guk; Lim, Jong Hyun; Park, Juhun; Hong, Seunghun; Ko, Hwi Jin; Park, Tai Hyun

    2015-12-15

    A bioelectronic nose for the real-time assessment of water quality was constructed with human olfactory receptor (hOR) and single-walled carbon nanotube field-effect transistor (swCNT-FET). Geosmin (GSM) and 2-methylisoborneol (MIB), mainly produced by bacteria, are representative odor compounds and also indicators of contamination in the water supply system. For the screening of hORs which respond to these compounds, we performed CRE-luciferase assays of the two odorants in heterologous cell system. Human OR51S1 for GSM and OR3A4 for MIB were selected, and nanovesicles expressing the hORs on surface were produced from HEK-293 cell. Carbon nanotube field-effect transistor was functionalized with the nanovesicles. The bioelectronic nose was able to selectively detect GSM and MIB at concentrations as low as a 10 ng L(-1). Furthermore, detection of these compounds from the real samples such as tap water, bottled water and river water was available without any pretreatment processes.

  2. Therapeutic and cosmeceutical potential of ethosomes: An overview

    Directory of Open Access Journals (Sweden)

    Poonam Verma

    2010-01-01

    Full Text Available The main disadvantage of transdermal drug delivery is the poor penetration of most compounds into the human skin. The main barrier of the skin is located within its uppermost layer, the stratum corneum (SC. Several approaches have been developed to weaken this skin barrier. One of the approaches for increasing the skin penetration of drugs and many cosmetic chemicals is the use of vesicular systems, such as, liposomes and ethosomes. Ethosomes are phospholipid-based elastic nanovesicles containing a high content of ethanol (20-45%. Ethanol is known as an efficient permeation enhancer and has been added in the vesicular systems to prepare elastic nanovesicles. It can interact with the polar head group region of the lipid molecules, resulting in the reduction of the melting point of the stratum corneum lipid, thereby increasing lipid fluidity and cell membrane permeability. The high flexibility of vesicular membranes from the added ethanol permits the elastic vesicles to squeeze themselves through the pores, which are much smaller than their diameters. Ethosomal systems are much more efficient in delivering substances to the skin in the terms of quantity and depth, than either conventional liposomes or hydroalcoholic solutions. The scope of this small review is to introduce the novel concept of ethosomes and to describe some approaches and mechanisms of stimulating topical and transdermal products with ethosomes.

  3. Therapeutic and cosmeceutical potential of ethosomes: An overview.

    Science.gov (United States)

    Verma, Poonam; Pathak, K

    2010-07-01

    The main disadvantage of transdermal drug delivery is the poor penetration of most compounds into the human skin. The main barrier of the skin is located within its uppermost layer, the stratum corneum (SC). Several approaches have been developed to weaken this skin barrier. One of the approaches for increasing the skin penetration of drugs and many cosmetic chemicals is the use of vesicular systems, such as, liposomes and ethosomes. Ethosomes are phospholipid-based elastic nanovesicles containing a high content of ethanol (20-45%). Ethanol is known as an efficient permeation enhancer and has been added in the vesicular systems to prepare elastic nanovesicles. It can interact with the polar head group region of the lipid molecules, resulting in the reduction of the melting point of the stratum corneum lipid, thereby increasing lipid fluidity and cell membrane permeability. The high flexibility of vesicular membranes from the added ethanol permits the elastic vesicles to squeeze themselves through the pores, which are much smaller than their diameters. Ethosomal systems are much more efficient in delivering substances to the skin in the terms of quantity and depth, than either conventional liposomes or hydroalcoholic solutions. The scope of this small review is to introduce the novel concept of ethosomes and to describe some approaches and mechanisms of stimulating topical and transdermal products with ethosomes.

  4. RGD-conjugated albumin nanoparticles as a novel delivery vehicle in pancreatic cancer therapy.

    Science.gov (United States)

    Ji, Shunrong; Xu, Jin; Zhang, Bo; Yao, Wantong; Xu, Wenyan; Wu, Wenzhe; Xu, Yongfeng; Wang, Hao; Ni, Quanxing; Hou, Huimin; Yu, Xianjun

    2012-02-15

    Integrin αvβ3 receptor is expressed on several types of cancer cells, including pancreatic cancer cells, and plays an important role in tumor growth and metastasis. The ability to target the integrin αvβ3 receptor on cancer cells increases the efficacy of targeted therapy and reduces the side effects. The aim of this study is to develop a novel arginine-glycine-aspartic acid (RGD) peptide -conjugated albumin nanoparticle to enhance the intracellular uptake of anticancer drug into the pancreatic cancer cells through receptor-mediated endocytosis. In the cellular uptake studies, the fluorescent signal of RGD-conjugated BSANPs in BxPC3 cells was higher than that of BSANPs without RGD conjugation as determined by fluorescence spectrophotometer. We also found that BSANPs bound to BxPC3 cells in a time- and concentration-dependent manner. The uptake of RGD-conjugated BSANPs by pancreatic cancer cells was inhibited by an excess amount of free RGD peptide, indicating that the binding and/or uptake were mediated by the αvβ3 receptor. Furthermore, the nanoparticles were found to be located close to the nuclei by using laser scanning confocal microscopy. Besides, no significant in vitro cytotoxicity was observed as measured with MTT assay. Both in vitro and in vivo antitumor efficacy was improved by targeting gemcitabine-loaded nanoparticles to BxPC-3 cells using RGD peptides. Therefore, the RGD-conjugated BSANPs hold great potential as an effective drug delivery system to deliver therapeutic agents to pancreatic cancer.

  5. Investigation of Porphyrin and Lipid Supramolecular Assemblies for Cancer Imaging and Therapy

    Science.gov (United States)

    Ng, Kenneth Ka-Seng

    Aerobic life on earth is made possible through the functions of the porphyrin. These colorful and ubiquitous chromophores are efficient at concentrating and converting sunlight into chemical energetic potential which sustain biological life. Humans have had a longstanding fascination with these molecules, especially for their applications in photodynamic therapy. The photophysical properties of porphyrins are highly influenced by their surrounding environment. Intermolecular interactions between these pigments can lead to excited state quenching, energy transfer and large changes to their absorption and fluorescence spectra. This thesis is focused on utilizing molecular self-assembly strategies to develop nanoscale porphyrin and phospholipid structures. The rationale being that intermolecular interactions between porphyrins in these nanostructures can induce changes which can be exploited in novel biomedical imaging and therapeutic applications. Four lipid-based structural platforms are studied including: nanoemulsions, bilayer discs and nanovesicles. In Chapter 1, I provide a background on the photophysics of porphyrins and the effect of intermolecular porphyrin interactions on photophysical properties. I also discuss phospholipids and their self-assembly process. Lastly I review current biomedical photonics techniques and discuss how these strategies can be used in conjugation with porphyrin and lipid supramolecular assemblies. In Chapter 2, I investigate the influence that loading a novel bacteriochlorin photosensitizer into a protein-stabilized lipid emulsion has on its spectral properties. I discovered that while the dye can be incorporated into the lipid emulsion, no changes were observed in its spectral properties. In Chapter 3, an amphipathic alpha-helical protein is used to stabilize and organize porphyrin-lipid molecules into bilayer discs. Close packing between porphyrin molecules causes quenching, which can be reversed by structural degradation of the

  6. Less is more: removing membrane attachments stiffens the RBC cytoskeleton

    Energy Technology Data Exchange (ETDEWEB)

    Gov, Nir S [Department of Chemical Physics, The Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel)

    2007-11-15

    The polymerized network of the cytoskeleton of the red-blood cell (RBC) contains different protein components that maintain its overall integrity and attachment to the lipid bilayer. One of these key components is the band 3-ankyrin complex that attaches the spectrin filaments to the fluid bilayer. Defects in this particular component result in the shape transformation called spherocytosis, through the shedding of membrane nano-vesicles. We show here that this transition and membrane shedding can be explained through the increased stiffness of the network when the band 3-ankyrin complexes are removed. ATP-induced transient dissociations lead to network softening, which offsets the stiffening to some extent, and causes increased fragility of these mutant cells, as is observed.

  7. Organized Aggregation of Porphyrins in Lipid Bilayers for Third Harmonic Generation Microscopy.

    Science.gov (United States)

    Cui, Liyang; Tokarz, Danielle; Cisek, Richard; Ng, Kenneth K; Wang, Fan; Chen, Juan; Barzda, Virginijus; Zheng, Gang

    2015-11-16

    Nonlinear optical microscopy has become a powerful tool for high-resolution imaging of cellular and subcellular composition, morphology, and interactions because of its high spatial resolution, deep penetration, and low photo-damage to tissue. Developing specific harmonic probes is essential for exploiting nonlinear microscopic imaging for biomedical applications. We report an organized aggregate of porphyrins (OAP) that formed within lipidic nanoparticles showing fingerprint spectroscopic properties, structure-associated second harmonic generation, and superradiant third harmonic generation. The OAP facilitated harmonic microscopic imaging of living cells with significantly enhanced contrast. The structure-dependent switch between harmonic (OAP-intact) and fluorescence (OAP-disrupted) generation enabled real-time multi-modality imaging of the cellular fate of nanoparticles. Robustly produced under various conditions and easily incorporated into pre-formed lipid nanovesicles, OAP provides a biocompatible nanoplatform for harmonic imaging. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Argan oil nanoemulsions as new hydrophobic drug-loaded delivery system for transdermal application.

    Science.gov (United States)

    Lococo, D; Mora-Huertas, C E; Fessi, H; Zaanoun, I; Elaissari, A

    2012-10-01

    This research work deals with the development of argan oil-based nanoemulsions as vehicle of hydrophobic drugs such as diclofenac used as model. Nanoemulsions of oil in water were prepared using the ultrasonication method in order to obtain submicron size colloidal dispersion. The size, zeta potential and encapsulation efficiency of the dispersions obtained were investigated. In addition, the ability of sorbitan ester derivatives to form nanovesicles (niosomes), which in turn were used for encapsulating drug in oily solutions forming stable nanoemulsions, was particularly examined. Thus, additional stabilizing agents were not required in the recipe and formulations using only sorbitan monolaurate, argan oil and water lead to attractive results. Their submicronic size (<250 nm), high negative zeta potential (between -40 and -50 mV) and drug-encapsulation efficiency (higher than 85%) allow predicting both a good physical stability and a good performance as drug carriers.

  9. Proteomic characterization of macro-, micro- and nano-extracellular vesicles derived from the same first trimester placenta: relevance for feto-maternal communication.

    Science.gov (United States)

    Tong, Mancy; Kleffmann, Torsten; Pradhan, Shantanu; Johansson, Caroline L; DeSousa, Joana; Stone, Peter R; James, Joanna L; Chen, Qi; Chamley, Larry W

    2016-04-01

    What proteins are carried by extracellular vesicles (EVs) released from normal first trimester placentae? One thousand five hundred and eighty-five, 1656 and 1476 proteins were characterized in macro-, micro- and nano-vesicles, respectively, from first trimester placentae, with all EV fractions being enriched for proteins involved in vesicle transport and inflammation. Placental EVs are being increasingly recognized as important mediators of both healthy and pathological pregnancies. However, current research has focused on detecting changes in specific proteins in particular fractions of vesicles during disease. This is the first study to investigate the full proteome of different-sized fractions of EVs from the same first trimester placenta and highlights the differences/similarities between the vesicle fractions. A well-established ex vivo placental explant culture model was used to generate macro-, micro- and nano-vesicles from 56 first trimester placentae. Vesicle fractions were collected by differential ultracentrifugation, quantified and characterized. Placental macro-, micro- and nano-vesicles were characterized by microscopy, dynamic light scattering and nanoparticle tracking analysis. The proteome of each EV fraction was interrogated using liquid chromatography-coupled tandem mass spectrometry. Results were validated by semi-quantitative western blotting. A total of 1585, 1656 and 1476 proteins were identified in macro-, micro- and nano-vesicles, respectively. One thousand one hundred and twenty-five proteins were shared between all three fractions while up to 223 proteins were unique to each fraction. Gene Ontology pathway analysis showed an enrichment of proteins involved in vesicle transport and inflammation in all three fractions of EVs. The expression levels of proteins involved in internalization of vesicles (annexin V, calreticulin, CD31, CD47), the complement pathway [C3, decay-accelerating factor (DAF), membrane cofactor protein (MCP), protectin

  10. Placental extracellular vesicles and feto-maternal communication.

    Science.gov (United States)

    Tong, M; Chamley, L W

    2015-01-29

    The human placenta is an anatomically unique structure that extrudes a variety of extracellular vesicles into the maternal blood (including syncytial nuclear aggregates, microvesicles, and nanovesicles). Large quantities of extracellular vesicles are produced by the placenta in both healthy and diseased pregnancies. Since their first description more than 120 years ago, placental extracellular vesicles are only now being recognized as important carriers for proteins, lipids, and nucleic acids, which may play a crucial role in feto-maternal communication. Here, we summarize the current literature on the cargos of placental extracellular vesicles and the known effects of such vesicles on maternal cells/systems, especially those of the maternal immune and vascular systems. Copyright © 2015 Cold Spring Harbor Laboratory Press; all rights reserved.

  11. Exosomes in human semen restrict HIV-1 transmission by vaginal cells and block intravaginal replication of LP-BM5 murine AIDS virus complex.

    Science.gov (United States)

    Madison, Marisa N; Jones, Philip H; Okeoma, Chioma M

    2015-08-01

    Exosomes are membranous extracellular nanovesicles secreted by diverse cell types. Exosomes from healthy human semen have been shown to inhibit HIV-1 replication and to impair progeny virus infectivity. In this study, we examined the ability of healthy human semen exosomes to restrict HIV-1 and LP-BM5 murine AIDS virus transmission in three different model systems. We show that vaginal cells internalize exosomes with concomitant transfer of functional mRNA. Semen exosomes blocked the spread of HIV-1 from vaginal epithelial cells to target cells in our cell-to-cell infection model and suppressed transmission of HIV-1 across the vaginal epithelial barrier in our trans-well model. Our in vivo model shows that human semen exosomes restrict intravaginal transmission and propagation of murine AIDS virus. Our study highlights an antiretroviral role for semen exosomes that may be harnessed for the development of novel therapeutic strategies to combat HIV-1 transmission.

  12. Physiological and pathological impact of exosomes of adipose tissue.

    Science.gov (United States)

    Zhang, Yan; Yu, Mei; Tian, Weidong

    2016-02-01

    Exosomes are nanovesicles that have emerged as a new intercellular communication system for transporting proteins and RNAs; recent studies have shown that they play a role in many physiological and pathological processes such as immune regulation, cell differentiation, infection and cancer. By transferring proteins, mRNAs and microRNAs, exosomes act as information vehicles that alter the behavior of recipient cells. Compared to direct cell-cell contact or secreted factors, exosomes can affect recipient cells in more efficient ways. In whole adipose tissues, it has been shown that exosomes exist in supernatants of adipocytes and adipose stromal cells (ADSCs). Adipocyte exosomes are linked to lipid metabolism and obesity-related insulin resistance and exosomes secreted by ADSCs are involved in angiogenesis, immunomodulation and tumor development. This review introduces characteristics of exosomes in adipose tissue, summarizes their functions in different physiological and pathological processes and provides the further insight into potential application of exosomes to disease diagnosis and treatment.

  13. Electrokinetic Evaluation of Individual Exosomes by On-Chip Microcapillary Electrophoresis with Laser Dark-Field Microscopy

    Science.gov (United States)

    Kato, Kei; Kobayashi, Masashi; Hanamura, Nami; Akagi, Takanori; Kosaka, Nobuyoshi; Ochiya, Takahiro; Ichiki, Takanori

    2013-06-01

    Cell-secreted nanovesicles called exosomes are expected as a promising candidate biomarker of various diseases. Toward the future application of exosomes as a disease biomarker for low-invasive diagnostics, challenges remain in the development of sensitive and precise analysis methods for exosomes. In this study, we performed the electrokinetic evaluation of individual exosomes by the combined use of on-chip microcapillary electrophoresis and laser dark-field microscopy. We extracted exosomes from six types of human cell cultured in a serum-free medium by differential ultracentrifugation and their zeta potential (electrophoretic mobility) were evaluated. We demonstrated that the proposed electrophoresis apparatus is particularly suitable for the tracking analysis of the electrophoretic migration of individual exosomes and enables the accurate evaluation of the zeta potential distribution of exosomes, for the first time. From the experimental results, we found that there is a strong correlation between the average zeta potentials of exosomes and their cells of origin.

  14. Texosome-based drug delivery system for cancer therapy:from past to present

    Institute of Scientific and Technical Information of China (English)

    Hamideh Mahmoodzadeh Hosseini; Raheleh Halabian; Mohsen Amin; Abbas Ali Imani Fooladi

    2015-01-01

    Rising worldwide cancer incidence and resistance to current anti-cancer drugs necessitate the need for new pharmaceutical compounds and drug delivery system. Malfunction of the immune system, particularly in the tumor microenvironment, causes tumor growth and enhances tumor progression. Thus, cancer immunotherapy can be an appropriate approach to provoke the systemic immune system to combat tumor expansion. Texosomes, which are endogenous nanovesicles released by all tumor cells, contribute to cell-cell communication and modify the phenotypic features of recipient cells due to the texosomes’ ability to transport biological components. For this reason, texosome-based delivery system can be a valuable strategy for therapeutic purposes. To improve the pharmaceutical behavior of this system and to facilitate its use in medical applications, biotechnology approaches and mimetic techniques have been utilized. In this review, we present the development history of texosome-based delivery systems and discuss the advantages and disadvantages of each system.

  15. Proteomic profiling of human plasma exosomes identifies PPARgamma as an exosome-associated protein.

    Science.gov (United States)

    Looze, Christopher; Yui, David; Leung, Lester; Ingham, Matthew; Kaler, Maryann; Yao, Xianglan; Wu, Wells W; Shen, Rong-Fong; Daniels, Mathew P; Levine, Stewart J

    2009-01-16

    Exosomes are nanovesicles that are released from cells as a mechanism of cell-free intercellular communication. Only a limited number of proteins have been identified from the plasma exosome proteome. Here, we developed a multi-step fractionation scheme incorporating gel exclusion chromatography, rate zonal centrifugation through continuous sucrose gradients, and high-speed centrifugation to purify exosomes from human plasma. Exosome-associated proteins were separated by SDS-PAGE and 66 proteins were identified by LC-MS/MS, which included both cellular and extracellular proteins. Furthermore, we identified and characterized peroxisome proliferator-activated receptor-gamma (PPARgamma), a nuclear receptor that regulates adipocyte differentiation and proliferation, as well as immune and inflammatory cell functions, as a novel component of plasma-derived exosomes. Given the important role of exosomes as intercellular messengers, the discovery of PPARgamma as a component of human plasma exosomes identifies a potential new pathway for the paracrine transfer of nuclear receptors.

  16. The human urinary exosome as a potential metabolic effector cargo.

    Science.gov (United States)

    Bruschi, Maurizio; Ravera, Silvia; Santucci, Laura; Candiano, Giovanni; Bartolucci, Martina; Calzia, Daniela; Lavarello, Chiara; Inglese, Elvira; Petretto, Andrea; Ghiggeri, Gianmarco; Panfoli, Isabella

    2015-08-01

    Exosomes are nanovesicles, derived from the endocytic pathway, released by most cell types and found in many body fluids, including urine. A variety of exosomal functions have been reported, including transfer of RNA, cell communication, control of apoptosis and protein lifespan. Exosomes from mesenchymal stem cells can rescue bioenergetics of injured cells. Here the urinary exosome proteome, non-urinary exosome proteome and urinome are compared. A consistent number of identified proteins cluster to metabolic functions. Cytoscape software analysis based on biological processes gene ontology database shows that metabolic pathways such as aerobic glycolysis and oxidative phosphorylation have a high probability (p ≤ 0.05) of being expressed and therefore functional. A metabolic function appears to be associated with human urinary exosomes, whose relevance experimental studies can assess.

  17. The Dual Role of Exosomes in Hepatitis A and C Virus Transmission and Viral Immune Activation.

    Science.gov (United States)

    Longatti, Andrea

    2015-12-17

    Exosomes are small nanovesicles of about 100 nm in diameter that act as intercellular messengers because they can shuttle RNA, proteins and lipids between different cells. Many studies have found that exosomes also play various roles in viral pathogenesis. Hepatitis A virus (HAV; a picornavirus) and Hepatitis C virus (HCV; a flavivirus) two single strand plus-sense RNA viruses, in particular, have been found to use exosomes for viral transmission thus evading antibody-mediated immune responses. Paradoxically, both viral exosomes can also be detected by plasmacytoid dendritic cells (pDCs) leading to innate immune activation and type I interferon production. This article will review recent findings regarding these two viruses and outline how exosomes are involved in their transmission and immune sensing.

  18. Exosome levels in human body fluids: A tumor marker by themselves?

    Science.gov (United States)

    Cappello, Francesco; Logozzi, Mariantonia; Campanella, Claudia; Bavisotto, Celeste Caruso; Marcilla, Antonio; Properzi, Francesca; Fais, Stefano

    2017-01-01

    Despite considerable research efforts, the finding of reliable tumor biomarkers remains challenging and unresolved. In recent years a novel diagnostic biomedical tool with high potential has been identified in extracellular nanovesicles or exosomes. They are released by the majority of the cells and contain detailed molecular information on the cell of origin including tumor hallmarks. Exosomes can be isolated from easy accessible body fluids, and most importantly, they can provide several biomarkers, with different levels of specificity. Recent clinical evidence shows that the levels of exosomes released into body fluids may themselves represent a predictive/diagnostic of tumors, discriminating cancer patients from healthy subjects. The aim of this review is to highlight these latest challenging findings to provide novel and groundbreaking ideas for successful tumor early diagnosis and follow-up.

  19. Exosomes: mediators of communication in eukaryotes.

    Science.gov (United States)

    Lopez-Verrilli, María A; Court, Felipe A

    2013-01-01

    In addition to the established mechanisms of intercellular signaling, a new way of communication has gained much attention in the last decade: communication mediated by exosomes. Exosomes are nanovesicles (with a diameter of 40-120 nm) secreted into the extracellular space by the multivesicular endosome after its outer membrane fuses with the plasma membrane. Once released, exosomes modulate the response of the recipient cells that recognize them. This indicates that exosomes operate in a specific manner and participate in the regulation of the target cell. Remarkably, exosomes occur from unicellular organisms to mammals, suggesting an evolutionarily conserved mechanism of communication. In this review we describe the cascade of exosome formation, intracellular traffic, secretion, and internalization by recipient cells, and review their most relevant effects. We also highlight important steps that are still poorly understood.

  20. Imaging and force measurement of LDL and HDL by AFM in air and liquid.

    Science.gov (United States)

    Gan, Chaoye; Ao, Meiying; Liu, Zhanghua; Chen, Yong

    2015-01-01

    The size and biomechanical properties of lipoproteins are tightly correlated with their structures/functions. While atomic force microscopy (AFM) has been used to image lipoproteins the force measurement of these nano-sized particles is missing. We detected that the sizes of LDL and HDL in liquid are close to the commonly known values. The Young's modulus of LDL or HDL is ∼0.4 GPa which is similar to that of some viral capsids or nanovesicles but greatly larger than that of various liposomes. The adhesive force of LDL or HDL is small (∼200 pN). The comparison of AFM detection in air and liquid was also performed which is currently lacking. Our data may provide useful information for better understanding and AFM detection of lipoproteins.

  1. Schwann cell-derived exosomes enhance axonal regeneration in the peripheral nervous system.

    Science.gov (United States)

    Lopez-Verrilli, María Alejandra; Picou, Frederic; Court, Felipe A

    2013-11-01

    Axonal regeneration in the peripheral nervous system is greatly supported by Schwann cells (SCs). After nerve injury, SCs dedifferentiate to a progenitor-like state and efficiently guide axons to their original target tissues. Contact and soluble factors participate in the crosstalk between SCs and axons during axonal regeneration. Here we show that dedifferentiated SCs secrete nano-vesicles known as exosomes which are specifically internalized by axons. Surprisingly, SC-derived exosomes markedly increase axonal regeneration in vitro and enhance regeneration after sciatic nerve injury in vivo. Exosomes shift the growth cone morphology to a pro-regenerating phenotype and decrease the activity of the GTPase RhoA, involved in growth cone collapse and axon retraction. Altogether, our work identifies a novel mechanism by which SCs communicate with neighboring axons during regenerative processes. We propose that SC exosomes represent an important mechanism by which these cells locally support axonal maintenance and regeneration after nerve damage.

  2. Comparative study of transfersomes, liposomes, and niosomes for topical delivery of 5-fluorouracil to skin cancer cells: preparation, characterization, in-vitro release, and cytotoxicity analysis.

    Science.gov (United States)

    Alvi, Iqrar Ali; Madan, Jitender; Kaushik, Dinesh; Sardana, Satish; Pandey, Ravi Shankar; Ali, Asgar

    2011-09-01

    Topical 5-fluorouracil (5-FU) is used for the treatment of actinic keratosis and nonmelanoma skin cancer. Unfortunately, 5-FU per se shows a poor percutaneous permeation, thus reducing its anticancer effectiveness after topical administration. Therefore, we have constructed transfersomes, liposomes, and niosomes of 5-FU for topical applications in this investigation. Transfersomes were prepared by the solvent evaporation method, whereas liposomes and niosomes were constructed by reverse-phase evaporation method. The nanovesicles were characterized for particle size, shape, zeta potential, viscosity, entrapment efficiency, deformability, in-vitro permeation release, and kinetics and retention. Cytotoxicity study was carried out on HaCaT cells. Transfersomes (153.2 ± 10.3 nm), liposomes (120.3 ± 9.8 nm), and niosomes (250.4 ± 8.6 nm) were produced with a maximum entrapment efficiency of 82.4 ± 4.8, 45.4 ± 3.3, and 43.4 ± 3.2%, respectively. Moreover, transmission electron microscopy and atomic force microscopy assure the smooth and spherical shape of nanovesicles. Skin permeation and retention showed better permeability and retention than the nonvesiculized dosage form. The IC50 value of transfersomes (1.02 μmol/l), liposomes (6.83 μmol/l), and niosomes (9.91 μmol/l) was found to be far less than 5-FU (15.89 μmol/l) at 72 h. 5-FU-loaded transfersomes were found to be most cytotoxic on the HaCaT cell line in comparison with liposomes and niosomes. We concluded that vesiculization of 5-FU not only improves the topical delivery, but also enhances the cytotoxic effect of 5-FU. We have presented here a viable formulation of 5-FU for the management of actinic keratosis and nonmelanoma skin carcinoma.

  3. Exosomes: The Link Between GPCR Activation and Metastatic Potential?

    Directory of Open Access Journals (Sweden)

    Allison Leigh Isola

    2016-04-01

    Full Text Available The activation of G-Protein Coupled Receptors (GPCRs by their respective ligands initiates a cascade of multiple signaling processes within the cell, regulating growth, metabolism and other essential cellular functions. Dysregulation and aberrant expression of these GPCRs and their subsequent signaling cascades are associated with many different types of pathologies, including cancer. The main life threatening complication in patients diagnosed with cancer is the dissemination of cells from the primary tumor to distant vital organs within the body, metastasis. Communication between the primary tumor, immune system, and the site of future metastasis are some of the key events in the early stages of metastasis. It has been postulated that the communication is mediated by nanovesicles that, under non-pathological conditions, are released by normal cells to relay signals to other cells in the body. These nanovesicles are called exosomes, and are utilized by the tumor cell to influence changes within the recipient cell, such as bone marrow progenitor cells, and cells within the site of future metastatic growth, in order to prepare the site for colonization. Tumor cells have been shown to release an increased number of exosomes when compared to their normal cell counterpart. Exosome production and release are regulated by proteins involved in localization, degradation and size of the multivesicular body, whose function may be altered within cancer cells, resulting in the release of an increased number of these vesicles. This review investigates the possibility of GPCR signaling cascades acting as the upstream activator of proteins involved in exosome production and release, linking a commonly targeted trans-membrane protein class with cellular communication utilized by tumor cells in early stages of metastasis.

  4. Identification and nanoentrapment of polyphenolic phytocomplex from Fraxinus angustifolia: in vitro and in vivo wound healing potential.

    Science.gov (United States)

    Moulaoui, Kenza; Caddeo, Carla; Manca, Maria Letizia; Castangia, Ines; Valenti, Donatella; Escribano, Elvira; Atmani, Djebbar; Fadda, Anna Maria; Manconi, Maria

    2015-01-07

    The aim of the present study was to elucidate the polyphenolic composition of Fraxinus angustifolia leaf and bark extracts, and to evaluate their efficacy in wound healing. Quercetin, catechin, rutin and tannic acid were identified as the main components of the extracts. In order to improve their skin bioavailability, the polyphenolic phytocomplexes were incorporated in different nanovesicles, namely ethosomes and phospholipid vesicles containing Transcutol(®) P (Trc) or ethylene glycol (EG). The latter had never been used before as a component of phospholipid vesicles, and it was found to play a key role in improving extract efficacy in wound healing. Results of cryogenic transmission electron microscopy (cryo-TEM), Photon Correlation Spectroscopy (PCS) and Small-Angle X-ray Scattering (SAXS) showed that ethosomes and EG-PEVs were small, monodispersed, unilamellar vesicles, while Trc-PEVs were larger, less homogeneously dispersed and multilamellar, with a large bilayer thickness. Free extracts did not show relevant ability to protect in vitro human keratinocytes from H2O2 damages, while when entrapped in nanovesicles, they significantly inhibited H2O2 stress damages, probably related to a higher level of cell uptake. On the other hand, in vivo results showed that the highest antioxidant and anti-inflammatory effects were provided by the phytocomplexes in EG-PEVs, which favoured wound healing. Moreover, non-entrapped F. angustifolia extracts showed a marginal effect, comparable to that of free quercetin dispersion (control). In conclusion, our results depict that these extracts may find potential applications in biomedicine.

  5. Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response

    Science.gov (United States)

    Menay, Florencia; Herschlik, Leticia; De Toro, Julieta; Cocozza, Federico; Tsacalian, Rodrigo; Gravisaco, María José; Di Sciullo, María Paula; Vendrell, Alejandrina; Waldner, Claudia I.; Mongini, Claudia

    2017-01-01

    Extracellular vesicles (EVs), including endosome-derived nanovesicles (exosomes), are involved in cell–cell communication. Through transfer of their molecular contents, extracellular nanovesicles can alter the function of recipient cells. Due to these characteristics, EVs have shown potential as a new alternative for cancer immunotherapy. Tumor exosomes isolated from malignant ascites can activate dendritic cells, thereby priming the immune system to recognize and kill cancer cells. However, a suppressive role on tumor immune response has also been reported, suggesting that the neoplastic stage of carcinogenesis and the microenvironment where tumor cells grow may influence the amount of EVs released by the cell. This neoplastic stage and microenvironment may also impact EVs’ components such as proteins and miRNA, determining their biological behavior. Most T-cell lymphomas have an aggressive clinical course and poor prognosis. Consequently, complementary alternative therapies are needed to improve the survival rates achieved with conventional treatments. In this work, we have characterized EVs isolated from ascites of mice bearing a very aggressive murine T-cell lymphoma and have studied their immunogenic properties. Small EVs were isolated by differential centrifugation, ultrafiltration, and ultracentrifugation at 100,000 × g on a sucrose cushion. The EVs were defined as exosomes by their morphology and size analyzed by electron microscopy, their floating density on a sucrose gradient, as well as their expression of endosome marker proteins ALIX, TSG-101; the tetraspanins CD63, CD9, and CD81. In addition, they contain tumor antigens, the marker for malignancy CD24, the heat shock protein HSP-70, and an unusual surface expression of HSP-90 was demonstrated. The administration of EVs isolated from ascites (EVs A) into naïve-syngeneic mice induced both humoral and cellular immune responses that allowed the rejection of subsequent tumor challenges. However

  6. Chlorosome-Inspired Synthesis of Templated Metallochlorin-Lipid Nanoassemblies for Biomedical Applications.

    Science.gov (United States)

    Ng, Kenneth K; Takada, Misa; Harmatys, Kara; Chen, Juan; Zheng, Gang

    2016-04-26

    Chlorosomes are vesicular light-harvesting organelles found in photosynthetic green sulfur bacteria. These organisms thrive in low photon flux environments due to the most efficient light-to-chemical energy conversion, promoted by a protein-less assembly of chlorin pigments. These assemblies possess collective absorption properties and can be adapted for contrast-enhanced bioimaging applications, where maximized light absorption in the near-infrared optical window is desired. Here, we report a strategy for tuning light absorption toward the near-infrared region by engineering a chlorosome-inspired assembly of synthetic metallochlorins in a biocompatible lipid scaffold. In a series of synthesized chlorin analogues, we discovered that lipid conjugation, central coordination of a zinc metal into the chlorin ring, and a 3(1)-methoxy substitution were critical for the formation of dye assemblies in lipid nanovesicles. The substitutions result in a specific optical shift, characterized by a bathochromically shifted (72 nm) Qy absorption band, along with an increase in absorbance and circular dichroism as the ratio of dye-conjugated lipid was increased. These alterations in optical spectra are indicative of the formation of delocalized excitons states across each molecular assembly. This strategy of tuning absorption by mimicking the structures found in photosynthetic organisms may spur new opportunities in the development of biophotonic contrast agents for medical applications.

  7. Fractionation of Exosomes and DNA using Size-Based Separation at the Nanoscale

    Science.gov (United States)

    Wunsch, Benjamin; Smith, Joshua; Wang, Chao; Gifford, Stacey; Brink, Markus; Bruce, Robert; Solovitzky, Gustavo; Austin, Robert; Astier, Yann

    Exosomes, a key target of ``liquid biopsies'', are nano-vesicles found in nearly all biological fluids. Exosomes are secreted by eukaryotic and prokaryotic cells alike, and contain information about their originating cells, including surface proteins, cytoplasmic proteins, and nucleic acids. One challenge in studying exosome morphology is the difficulty of sorting exosomes by size and surface markers. Common separation techniques for exosomes include ultracentrifugation and ultrafiltration, for preparation of large volume samples, but these techniques often show contamination and significant heterogeneity between preparations. To date, deterministic lateral displacement (DLD) pillar arrays in silicon have proven an efficient technology to sort, separate, and enrich micron-scale particles including human parasites, eukaryotic cells, blood cells, and circulating tumor cells in blood; however, the DLD technology has never been translated to the true nanoscale, where it could function on bio-colloids such as exosomes. We have fabricated nanoscale DLD (nanoDLD) arrays capable of rapidly sorting colloids down to 20 nm in continuous flow, and demonstrated size sorting of individual exosome vesicles and dsDNA polymers, opening the potential for on-chip biomolecule separation and diagnosti

  8. Disposable pencil graphite electrode modified with peptide nanotubes for Vitamin B{sub 12} analysis

    Energy Technology Data Exchange (ETDEWEB)

    Pala, Betül Bozdoğan [Nanotechnology and Nanomedicine Division, Institute of Science, Hacettepe University, 06800 Ankara (Turkey); Vural, Tayfun [Department of Chemistry, Faculty of Science, Hacettepe University, 06800 Beytepe, Ankara (Turkey); Kuralay, Filiz [Department of Chemistry, Faculty of Science and Arts, Ordu University, 52200 Ordu (Turkey); Çırak, Tamer [Nanotechnology and Nanomedicine Division, Institute of Science, Hacettepe University, 06800 Ankara (Turkey); Bolat, Gülçin; Abacı, Serdar [Department of Chemistry, Faculty of Science, Hacettepe University, 06800 Beytepe, Ankara (Turkey); Denkbaş, Emir Baki, E-mail: denkbas@hacettepe.edu.tr [Department of Chemistry, Faculty of Science, Hacettepe University, 06800 Beytepe, Ankara (Turkey)

    2014-06-01

    In this study, peptide nanostructures from diphenylalanine were synthesized in various solvents with various polarities and characterized with Scanning Electron Microscopy (SEM) and Powder X-ray Diffraction (PXRD) techniques. Formation of peptide nanofibrils, nanovesicles, nanoribbons, and nanotubes was observed in different solvent mediums. In order to investigate the effects of peptide nanotubes (PNT) on electrochemical behavior of disposable pencil graphite electrodes (PGE), electrode surfaces were modified with fabricated peptide nanotubes. Electrochemical activity of the pencil graphite electrode was increased with the deposition of PNTs on the surface. The effects of the solvent type, the peptide nanotube concentration, and the passive adsorption time of peptide nanotubes on pencil graphite electrode were studied. For further electrochemical studies, electrodes were modified for 30 min by immobilizing PNTs, which were prepared in water at 6 mg/mL concentration. Vitamin B{sub 12} analyses were performed by the Square Wave (SW) voltammetry method using modified PGEs. The obtained data showed linearity over the range of 0.2 μM and 9.50 μM Vitamin B{sub 12} concentration with high sensitivity. Results showed that PNT modified PGEs were highly simple, fast, cost effective, and feasible for the electro-analytical determination of Vitamin B{sub 12} in real samples.

  9. Potential Therapies by Stem Cell-Derived Exosomes in CNS Diseases: Focusing on the Neurogenic Niche

    Directory of Open Access Journals (Sweden)

    Alejandro Luarte

    2016-01-01

    Full Text Available Neurodegenerative disorders are one of the leading causes of death and disability and one of the biggest burdens on health care systems. Novel approaches using various types of stem cells have been proposed to treat common neurodegenerative disorders such as Alzheimer’s Disease, Parkinson’s Disease, or stroke. Moreover, as the secretome of these cells appears to be of greater benefit compared to the cells themselves, the extracellular components responsible for its therapeutic benefit have been explored. Stem cells, as well as most cells, release extracellular vesicles such as exosomes, which are nanovesicles able to target specific cell types and thus to modify their function by delivering proteins, lipids, and nucleic acids. Exosomes have recently been tested in vivo and in vitro as therapeutic conveyors for the treatment of diseases. As such, they could be engineered to target specific populations of cells within the CNS. Considering the fact that many degenerative brain diseases have an impact on adult neurogenesis, we discuss how the modulation of the adult neurogenic niches may be a therapeutic target of stem cell-derived exosomes. These novel approaches should be examined in cellular and animal models to provide better, more effective, and specific therapeutic tools in the future.

  10. Urinary Exosomes

    Directory of Open Access Journals (Sweden)

    Irena Dimov

    2009-01-01

    Full Text Available Exosomes are nanovesicles of endocytic origin that are secreted into the extracellular space or body fluids when a multivesicular body (MVB fuses with the cell membrane. Interest in exosomes intensified after their description in antigen-presenting cells and the observation that they can significantly moderate immune responses in vivo. In the past few years, several groups have reported on the secretion of exosomes by almost all cell types in an organism. In addition to a common set of membrane and cytosolic molecules, exosomes harbor unique subsets of proteins, reflecting their cellular source. Major research efforts were put into their surprisingly various biological functions and in translating knowledge into clinical practice. Urine provides an exciting noninvasive alternative to blood or tissue samples as a potential source of disease biomarkers. Urinary exosomes (UE became the subject of serious studies just a few years ago. A recent large-scale proteomics-based study of normal UE revealed a myriad of proteins, including disease-related gene products. Thus, UE have valuable potential as a source of biomarkers for early detection of various types of diseases, monitoring the disease evolution and/or response to therapy. As a relatively new field of research, it still faces many challenges, but UE have already shown some straightforward potential.

  11. The bone marrow microenvironment enhances multiple myeloma progression by exosome-mediated activation of myeloid-derived suppressor cells.

    Science.gov (United States)

    Wang, Jinheng; De Veirman, Kim; De Beule, Nathan; Maes, Ken; De Bruyne, Elke; Van Valckenborgh, Els; Vanderkerken, Karin; Menu, Eline

    2015-12-22

    Exosomes, extracellular nanovesicles secreted by various cell types, modulate the bone marrow (BM) microenvironment by regulating angiogenesis, cytokine release, immune response, inflammation, and metastasis. Interactions between bone marrow stromal cells (BMSCs) and multiple myeloma (MM) cells play crucial roles in MM development. We previously reported that BMSC-derived exosomes directly promote MM cell growth, whereas the other possible mechanisms for supporting MM progression by these exosomes are still not clear. Here, we investigated the effect of BMSC-derived exosomes on the MM BM cells with specific emphasis on myeloid-derived suppressor cells (MDSCs). BMSC-derived exosomes were able to be taken up by MM MDSCs and induced their expansion in vitro. Moreover, these exosomes directly induced the survival of MDSCs through activating STAT3 and STAT1 pathways and increasing the anti-apoptotic proteins Bcl-xL and Mcl-1. Inhibition of these pathways blocked the enhancement of MDSC survival. Furthermore, these exosomes increased the nitric oxide release from MM MDSCs and enhanced their suppressive activity on T cells. Taken together, our results demonstrate that BMSC-derived exosomes activate MDSCs in the BM through STAT3 and STAT1 pathways, leading to increased immunosuppression which favors MM progression.

  12. Circulating exosomes as new biomarkers for brain disease and injury

    Science.gov (United States)

    Graner, Michael W.; Epple, Laura M.; Dusto, Nathaniel L.; Lencioni, Alex M.; Nega, Meheret; Herring, Matthew; Winston, Ben; Madsen, Helen; Bemis, Lynne T.; Anchordoquy, Thomas J.

    2013-05-01

    Brain diseases such as cancers, neurodegenerative disorders, or trauma are frequently diagnosed with imaging modalities and sometimes with intracranial biopsies. Treatment response is similarly monitored, along with clinical indications. While these technologies provide important windows into the disease state, they fail to provide us a detailed molecular portrait of the disease and of the changes taking place during therapy. Exosomes are virus-sized nanovesicles derived from the endosomal system and are released extracellularly from essentially all cell types. Exosomes contain intracellular entities (proteins, nucleic acids, metabolites), membrane proteins and lipids, and even extracellular proteins bound to them. Exosomes may be considered as mini-surrogates of their cells of origin, with some content common to all cells/exosomes, but some of the content would be cell-specific. These vesicles are found in all biofluids in humans, and are thus accessible to "liquid biopsy" with harvest of vesicles from such fluids. Current challenges are to identify disease-related markers or panels of markers to distinguish the disease state. Here we will show examples of brain tumor markers found in/on exosomes from cell culture and patient sera, and we will suggest that aspects of the biology of disease may have a relevant place in the search for biomarkers.

  13. Staphylococcus aureus produces membrane-derived vesicles that induce host cell death.

    Directory of Open Access Journals (Sweden)

    Mamata Gurung

    Full Text Available Gram-negative bacteria produce outer membrane vesicles that play a role in the delivery of virulence factors to host cells. However, little is known about the membrane-derived vesicles (MVs produced by gram-positive bacteria. The present study examined the production of MVs from Staphylococcus aureus and investigated the delivery of MVs to host cells and subsequent cytotoxicity. Four S. aureus strains tested, two type strains and two clinical isolates, produced spherical nanovesicles during in vitro culture. MVs were also produced during in vivo infection of a clinical S. aureus isolate in a mouse pneumonia model. Proteomic analysis showed that 143 different proteins were identified in the S. aureus-derived MVs. S. aureus MVs were interacted with the plasma membrane of host cells via a cholesterol-rich membrane microdomain and then delivered their component protein A to host cells within 30 min. Intact S. aureus MVs induced apoptosis of HEp-2 cells in a dose-dependent manner, whereas lysed MVs neither delivered their component into the cytosol of host cells nor induced cytotoxicity. In conclusion, this study is the first report that S. aureus MVs are an important vehicle for delivery of bacterial effector molecules to host cells.

  14. Development of exosome surface display technology in living human cells

    Energy Technology Data Exchange (ETDEWEB)

    Stickney, Zachary, E-mail: zstickney@scu.edu; Losacco, Joseph, E-mail: jlosacco@scu.edu; McDevitt, Sophie, E-mail: smmcdevitt@scu.edu; Zhang, Zhiwen, E-mail: zzhang@scu.edu; Lu, Biao, E-mail: blu2@scu.edu

    2016-03-25

    Surface display technology is an emerging key player in presenting functional proteins for targeted drug delivery and therapy. Although a number of technologies exist, a desirable mammalian surface display system is lacking. Exosomes are extracellular vesicles that facilitate cell–cell communication and can be engineered as nano-shuttles for cell-specific delivery. In this study, we report the development of a novel exosome surface display technology by exploiting mammalian cell secreted nano-vesicles and their trans-membrane protein tetraspanins. By constructing a set of fluorescent reporters for both the inner and outer surface display on exosomes at two selected sites of tetraspanins, we demonstrated the successful exosomal display via gene transfection and monitoring fluorescence in vivo. We subsequently validated our system by demonstrating the expected intracellular partitioning of reporter protein into sub-cellular compartments and secretion of exosomes from human HEK293 cells. Lastly, we established the stable engineered cells to harness the ability of this robust system for continuous production, secretion, and uptake of displayed exosomes with minimal impact on human cell biology. In sum, our work paved the way for potential applications of exosome, including exosome tracking and imaging, targeted drug delivery, as well as exosome-mediated vaccine and therapy.

  15. A Polydiacetylene Vesicle-based Colorimetric Detection of Campylobacter jejuni Using a DNA Aptamer as Recognition Element%聚丁二炔纳米囊泡生物传感器比色检测空肠弯曲菌的初步研究

    Institute of Scientific and Technical Information of China (English)

    董建涛; 欧阳后先; 徐柯; 吴文鹤; 吕建新

    2012-01-01

    A polydiacetylene vesicle-based biosensor for rapid colorimetric detection of Campylobacter jejuni was developed in this work. Aptamers coupled to the surface of polydiacetylene nanovesicle by chemical reaction in order to achieve vesicle functionalization. Molecular recognition between Campylobacter jejuni and aptamers on the surface of the vesicle lead to absorption spectrum transition of polydiacetylene by which quantitative detection of Campylobacter jejuni could be realized. This method can detect Campylobacter jejuni in 30 mins with high specificity and simple operation.%本实验构建一种可以快速比色检测空肠弯曲菌的聚丁二炔纳米囊泡生物传感器.适配体通过化学反应耦合到聚丁二炔纳米囊泡表面,完成对囊泡的功能化.适配体与空肠弯曲菌特异识别,引起纳米囊泡溶液吸收光谱的改变,通过比色响应值衡量检测体系中有无空肠弯曲菌.该方法特异性好,与PCR对比实验相符;操作简便,检测时间短,仅需30 min.

  16. Disposable pencil graphite electrode modified with peptide nanotubes for Vitamin B12 analysis

    Science.gov (United States)

    Pala, Betül Bozdoğan; Vural, Tayfun; Kuralay, Filiz; Çırak, Tamer; Bolat, Gülçin; Abacı, Serdar; Denkbaş, Emir Baki

    2014-06-01

    In this study, peptide nanostructures from diphenylalanine were synthesized in various solvents with various polarities and characterized with Scanning Electron Microscopy (SEM) and Powder X-ray Diffraction (PXRD) techniques. Formation of peptide nanofibrils, nanovesicles, nanoribbons, and nanotubes was observed in different solvent mediums. In order to investigate the effects of peptide nanotubes (PNT) on electrochemical behavior of disposable pencil graphite electrodes (PGE), electrode surfaces were modified with fabricated peptide nanotubes. Electrochemical activity of the pencil graphite electrode was increased with the deposition of PNTs on the surface. The effects of the solvent type, the peptide nanotube concentration, and the passive adsorption time of peptide nanotubes on pencil graphite electrode were studied. For further electrochemical studies, electrodes were modified for 30 min by immobilizing PNTs, which were prepared in water at 6 mg/mL concentration. Vitamin B12 analyses were performed by the Square Wave (SW) voltammetry method using modified PGEs. The obtained data showed linearity over the range of 0.2 μM and 9.50 μM Vitamin B12 concentration with high sensitivity. Results showed that PNT modified PGEs were highly simple, fast, cost effective, and feasible for the electro-analytical determination of Vitamin B12 in real samples.

  17. Stable J-aggregation enabled dual photoacoustic and fluorescence nanoparticles for intraoperative cancer imaging.

    Science.gov (United States)

    Shakiba, Mojdeh; Ng, Kenneth K; Huynh, Elizabeth; Chan, Harley; Charron, Danielle M; Chen, Juan; Muhanna, Nidal; Foster, F Stuart; Wilson, Brian C; Zheng, Gang

    2016-07-07

    J-aggregates display nanoscale optical properties which enable their use in fluorescence and photoacoustic imaging applications. However, control over their optical properties in an in vivo setting is hampered by the conformational lability of the J-aggregate structure in complex biological environments. J-aggregating nanoparticles (JNP) formed by self-assembly of bacteriopheophorbide-lipid (Bchl-lipid) in lipid nanovesicles represents a novel strategy to stabilize J-aggregates for in vivo bioimaging applications. We find that 15 mol% Bchl-lipid embedded within a saturated phospholipid bilayer vesicle was optimal in terms of maximizing Bchl-lipid dye loading, while maintaining a spherical nanoparticle morphology and retaining spectral properties characteristic of J-aggregates. The addition of cholesterol maintains the stability of the J-aggregate absorption band for up to 6 hours in the presence of 90% FBS. In a proof-of-concept experiment, we successfully applied JNPs as a fluorescence contrast agent for real-time intraoperative detection of metastatic lymph nodes in a rabbit head-and-neck cancer model. Lymph node metastasis delineation was further verified by visualizing the JNP within the excised lymph node using photoacoustic imaging. Using JNPs, we demonstrate the possibility of using J-aggregates as fluorescence and photoacoustic contrast agents and may potentially spur the development of other nanomaterials that can stably induce J-aggregation for in vivo cancer bioimaging applications.

  18. Impact of model perfume molecules on the self-assembly of anionic surfactant sodium dodecyl 6-benzene sulfonate.

    Science.gov (United States)

    Bradbury, Robert; Penfold, Jeffrey; Thomas, Robert K; Tucker, Ian M; Petkov, Jordan T; Jones, Craig; Grillo, Isabelle

    2013-03-12

    The impact of two model perfumes with differing degrees of hydrophobicity/hydrophilicity, linalool (LL) and phenylethanol (PE), on the solution structure of anionic surfactant sodium dodecyl 6-benzene sulfonate, LAS-6, has been studied by small angle neutron scattering, SANS. For both types of perfume molecules, complex phase behavior is observed. The phase behavior depends upon the concentration, surfactant/perfume composition, and type of perfume. The more hydrophilic perfume PE promotes the formation of more highly curved structures. At relatively low surfactant concentrations, small globular micelles, L1, are formed. These become perfume droplets, L(sm), stabilized by the surfactant at much higher perfume solution compositions. At higher surfactant concentrations, the tendency of LAS-6 to form more planar structures is evident. The more hydrophobic linalool promotes the formation of more planar structures. Combined with the greater tendency of LAS-6 to form planar structures, this results in the planar structures dominating the phase behavior for the LAS-6/linalool mixtures. For the LAS-6/linalool mixture, the self-assembly is in the form of micelles only at the lowest surfactant and perfume concentrations. Over most of the concentration-composition space explored, the structures are predominantly lamellar, L(α), or vesicle, L(v), or in the form of a lamellar/micellar coexistence. At low and intermediate amounts of LL, a significantly different structure is observed, and the aggregates are in the form of small, relatively monodisperse vesicles (i.e., nanovesicles), L(sv).

  19. Extracellular vesicles: fundamentals and clinical relevance

    Directory of Open Access Journals (Sweden)

    Wael Nassar

    2015-01-01

    Full Text Available All types of cells of eukaryotic organisms produce and release small nanovesicles into their extracellular environment. Early studies have described these vesicles as ′garbage bags′ only to remove obsolete cellular molecules. Valadi and colleagues, in 2007, were the first to discover the capability of circulating extracellular vesicles (EVs to horizontally transfer functioning gene information between cells. These extracellular vesicles express components responsible for angiogenesis promotion, stromal remodeling, chemoresistance, genetic exchange, and signaling pathway activation through growth factor/receptor transfer. EVs represent an important mode of intercellular communication by serving as vehicles for transfer between cells of membrane and cytosolic proteins, lipids, signaling proteins, and RNAs. They contribute to physiology and pathology, and they have a myriad of potential clinical applications in health and disease. Moreover, vesicles can pass the blood-brain barrier and may perhaps even be considered as naturally occurring liposomes. These cell-derived EVs not only represent a central mediator of the disease microenvironment, but their presence in the peripheral circulation may serve as a surrogate for disease biopsies, enabling real-time diagnosis and disease monitoring. In this review, we′ll be addressing the characteristics of different types of extracellular EVs, as well as their clinical relevance and potential as diagnostic markers, and also define therapeutic options.

  20. Exosomes: the ideal nanovectors for biodelivery.

    Science.gov (United States)

    Fais, Stefano; Logozzi, Mariantonia; Lugini, Luana; Federici, Cristina; Azzarito, Tommaso; Zarovni, Natasa; Chiesi, Antonio

    2013-01-01

    Nanomedicine aims to exploit the improved and often novel physical, chemical, and biological properties of materials at the nanometric scale, possibly with the highest level of biomimetism, an approach that simulates what occurs in nature. Although extracellularly released vesicles include both microvesicles (MVs) and exosomes, only exosomes have the size that may be considered suitable for potential use in nanomedicine. In fact, recent reports have shown that exosomes are able to interact with target cells within an organ or at a distance using different mechanisms. Much is yet to be understood about exosomes, and currently, we are looking at the visible top of an iceberg, with most of what we have to understand on these nanovesicles still under the sea. In fact, we know that exosomes released by normal cells always trigger positive effects, whereas those released by cells in pathological condition, such as tumor or infected cells, may induce undesired, dangerous, and mostly unknown effects, but we cannot exclude the possibility that exosomes may also be detrimental for the body in normal conditions. However, whether we consider extracellular vesicles as a whole, thus including MVs, it appears that even in normal conditions, extracellular vesicles may lead to unwanted effects, depending on gender and age. This review aims to critically emphasize existing data in the literature that support the possible roles of exosomes in both diagnostic and therapeutic scopes.

  1. Bovine milk-derived exosomes for drug delivery.

    Science.gov (United States)

    Munagala, Radha; Aqil, Farrukh; Jeyabalan, Jeyaprakash; Gupta, Ramesh C

    2016-02-01

    Exosomes are biological nanovesicles that are involved in cell-cell communication via the functionally-active cargo (such as miRNA, mRNA, DNA and proteins). Because of their nanosize, exosomes are explored as nanodevices for the development of new therapeutic applications. However, bulk, safe and cost-effective production of exosomes is not available. Here, we show that bovine milk can serve as a scalable source of exosomes that can act as a carrier for chemotherapeutic/chemopreventive agents. Drug-loaded exosomes showed significantly higher efficacy compared to free drug in cell culture studies and against lung tumor xenografts in vivo. Moreover, tumor targeting ligands such as folate increased cancer-cell targeting of the exosomes resulting in enhanced tumor reduction. Milk exosomes exhibited cross-species tolerance with no adverse immune and inflammatory response. Thus, we show the versatility of milk exosomes with respect to the cargo it can carry and ability to achieve tumor targetability. This is the first report to identify a biocompatible and cost-effective means of exosomes to enhance oral bioavailability, improve efficacy and safety of drugs.

  2. Vascular cemeteries formed by biological nanoparticles

    Science.gov (United States)

    Sommer, Andrei P.; Tsurumoto, Toshiyuki

    2013-04-01

    We report the discovery of dense colonies of globular structures ranging from 100 nm to 5 μm in the tunica media of the femoral artery of an 89-year-old female cadaver. Systematic analysis using scanning electron microscopy, energy dispersive X-ray spectroscopy and light microscopy reveals that the globular structures are surrounded by vascular smooth muscle cells (VSMCs) and consist predominantly of calcium phosphate. Inspection of the images suggests the action of two complementary growth processes. The structures may grow both in size and in number locally by Ostwald ripening and a replicative route, respectively. Morphology in conjunction with the quality of their native growth niche suggests that they are different from nanocrystals released from apoptotic bodies. Their tendency to fill VSMC pockets leads to the speculation that they could represent an effort of the VSMC system to wall off cytotoxic nanocrystals liberated from apoptotic bodies. Alternatively, the structures may be equivalent with nanobacteria (NB)—a nomenclature which caused confusion. This is reflected by the multitude of names used by different authors for the nanoentities (living nanovesicles, nanobionta, calcifying nanoparticles, and nanons). Indeed, there is no clear definition in the literature as to what NB are. Considering that the calcium phosphate nanoparticles have been identified in the human body, we used in our study the descriptive name biological nanoparticles—the world's first nanoparticles.

  3. Vascular cemeteries formed by biological nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Sommer, Andrei P., E-mail: andrei.sommer@uni-ulm.de [University of Ulm, Institute of Micro and Nanomaterials (Germany); Tsurumoto, Toshiyuki [Nagasaki University, Department of Macroscopic Anatomy, Graduate School of Biomedical Science (Japan)

    2013-04-15

    We report the discovery of dense colonies of globular structures ranging from 100 nm to 5 {mu}m in the tunica media of the femoral artery of an 89-year-old female cadaver. Systematic analysis using scanning electron microscopy, energy dispersive X-ray spectroscopy and light microscopy reveals that the globular structures are surrounded by vascular smooth muscle cells (VSMCs) and consist predominantly of calcium phosphate. Inspection of the images suggests the action of two complementary growth processes. The structures may grow both in size and in number locally by Ostwald ripening and a replicative route, respectively. Morphology in conjunction with the quality of their native growth niche suggests that they are different from nanocrystals released from apoptotic bodies. Their tendency to fill VSMC pockets leads to the speculation that they could represent an effort of the VSMC system to wall off cytotoxic nanocrystals liberated from apoptotic bodies. Alternatively, the structures may be equivalent with nanobacteria (NB)-a nomenclature which caused confusion. This is reflected by the multitude of names used by different authors for the nanoentities (living nanovesicles, nanobionta, calcifying nanoparticles, and nanons). Indeed, there is no clear definition in the literature as to what NB are. Considering that the calcium phosphate nanoparticles have been identified in the human body, we used in our study the descriptive name biological nanoparticles-the world's first nanoparticles.

  4. Tumor Microenvironment Modulation via Gold Nanoparticles Targeting Malicious Exosomes: Implications for Cancer Diagnostics and Therapy

    Science.gov (United States)

    Roma-Rodrigues, Catarina; Raposo, Luís R.; Cabral, Rita; Paradinha, Fabiana; Baptista, Pedro V.; Fernandes, Alexandra R.

    2017-01-01

    Exosomes are nanovesicles formed in the endosomal pathway with an important role in paracrine and autocrine cell communication. Exosomes secreted by cancer cells, malicious exosomes, have important roles in tumor microenvironment maturation and cancer progression. The knowledge of the role of exosomes in tumorigenesis prompted a new era in cancer diagnostics and therapy, taking advantage of the use of circulating exosomes as tumor biomarkers due to their stability in body fluids and targeting malignant exosomes’ release and/or uptake to inhibit or delay tumor development. In recent years, nanotechnology has paved the way for the development of a plethora of new diagnostic and therapeutic platforms, fostering theranostics. The unique physical and chemical properties of gold nanoparticles (AuNPs) make them suitable vehicles to pursuit this goal. AuNPs’ properties such as ease of synthesis with the desired shape and size, high surface:volume ratio, and the possibility of engineering their surface as desired, potentiate AuNPs’ role in nanotheranostics, allowing the use of the same formulation for exosome detection and restraining the effect of malicious exosomes in cancer progression. PMID:28098821

  5. Cross-linked chitosan/liposome hybrid system for the intestinal delivery of quercetin.

    Science.gov (United States)

    Caddeo, Carla; Díez-Sales, Octavio; Pons, Ramon; Carbone, Claudia; Ennas, Guido; Puglisi, Giovanni; Fadda, Anna Maria; Manconi, Maria

    2016-01-01

    Quercetin is a flavonoid with antioxidant/anti-inflammatory properties, poorly absorbed when administered orally. To increase its bioavailability and optimize its release in the intestine, a hybrid system made of liposomes coated with cross-linked chitosan, named TPP-chitosomes, was developed and characterized by light scattering, transmission electron microscopy, differential scanning calorimetry, X-ray powder diffraction and Turbiscan® technology. The TPP-chitosomes were nanosized (∼180 nm), fairly spherical in shape and unilamellar. The actual coating of the surface of liposomes with the cross-linked chitosan was demonstrated by Small-Angle X-ray Scattering. The release of quercetin in simulated gastric and intestinal pH was investigated, the results showing that the system provided resistance to acidic conditions, and promoted the release in alkaline pH, mimicking the intestinal environment. The proposed hybrid system represents a promising combination of nanovesicles and chitosan for the delivery of quercetin to the intestine in the therapy of oxidative stress/inflammation related disorders.

  6. Acronychiabaueri Analogue Derivative-Loaded Ultradeformable Vesicles: Physicochemical Characterization and Potential Applications.

    Science.gov (United States)

    Di Francesco, Martina; Primavera, Rosita; Fiorito, Serena; Cristiano, Maria Chiara; Taddeo, Vito Alessandro; Epifano, Francesco; Di Marzio, Luisa; Genovese, Salvatore; Celia, Christian

    2017-03-01

    Elastic and ultradeformable liposomes were synthesized and physicochemically characterized to make suitable topical formulations for delivering the anti-inflammatory and anticancer compound 3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans-propenoic acid. The average sizes of elastic and ultradeformable liposomes are below 300 nm, while the size distribution and Z-potential are below 0.3 and - 25 mV, respectively. The presence of 3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans-propenoic acid does not affect the physicochemical parameters of nanovesicles. Elastic and ultradeformable liposomes show a zero order release kinetic and are stable at room temperature for a long time with or without 3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans-propenoic acid. The ultradeformable liposomes are more deformable than elastic liposomes. These differences may depend on sodium cholate derivatives making nanoformulations. The 3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans-propenoic acid-loaded elastic and ultradeformable liposomes can provide innovative nanotherapeutics-based natural compounds for the potential treatment of cutanous inflammation.

  7. Analytical Applications of Nanomaterials in Monitoring Biological and Chemical Contaminants in Food.

    Science.gov (United States)

    Lim, Min-Cheol; Kim, Young-Rok

    2016-09-28

    The detection of food pathogens is an important aspect of food safety. A range of detection systems and new analytical materials have been developed to achieve fast, sensitive, and accurate monitoring of target pathogens. In this review, we summarize the characteristics of selected nanomaterials and their applications in food, and place focus on the monitoring of biological and chemical contaminants in food. The unique optical and electrical properties of nanomaterials, such as gold nanoparticles, nanorods, quantum dots, carbon nanotubes, graphenes, nanopores, and polydiacetylene nanovesicles, are closely associated with their dimensions, which are comparable in scale to those of targeted biomolecules. Furthermore, their optical and electrical properties are highly dependent on local environments, which make them promising materials for sensor development. The specificity and selectivity of analytical nanomaterials for target contaminants can be achieved by combining them with various biological entities, such as antibodies, oligonucleotides, aptamers, membrane proteins, and biological ligands. Examples of nanomaterial-based analytical systems are presented together with their limitations and associated developmental issues.

  8. Designer nanomaterials using chiral self-assembling peptide systems and their emerging benefit for society.

    Science.gov (United States)

    Luo, Zhongli; Zhang, Shuguang

    2012-07-07

    Chirality is absolutely central in chemistry and biology. The recent findings of chiral self-assembling peptides' remarkable chemical complementarity and structural compatibility make it one of the most inspired designer materials and structures in nanobiotechnology. The emerging field of designer chemistry and biology further explores biological and medical applications of these simple D,L- amino acids through producing marvellous nanostructures under physiological conditions. These self-assembled structures include well-ordered nanofibers, nanotubes and nanovesicles. These structures have been used for 3-dimensional tissue cultures of primary cells and stem cells, sustained release of small molecules, growth factors and monoclonal antibodies, accelerated wound-healing in reparative and regenerative medicine as well as tissue engineering. Recent advances in molecular designs have also led to the development of 3D fine-tuned bioactive tissue culture scaffolds. They are also used to stabilize membrane proteins including difficult G-protein coupled receptors for designing nanobiodevices. One of the self-assembling peptides has been used in human clinical trials for accelerated wound-healings. It is our hope that these peptide materials will open doors for more and diverse clinical uses. The field of chiral self-assembling peptide nanobiotechnology is growing in a number of directions that has led to many surprises in areas of novel materials, synthetic biology, clinical medicine and beyond.

  9. Potential Therapies by Stem Cell-Derived Exosomes in CNS Diseases: Focusing on the Neurogenic Niche

    Science.gov (United States)

    Luarte, Alejandro; Bátiz, Luis Federico; Wyneken, Ursula; Lafourcade, Carlos

    2016-01-01

    Neurodegenerative disorders are one of the leading causes of death and disability and one of the biggest burdens on health care systems. Novel approaches using various types of stem cells have been proposed to treat common neurodegenerative disorders such as Alzheimer's Disease, Parkinson's Disease, or stroke. Moreover, as the secretome of these cells appears to be of greater benefit compared to the cells themselves, the extracellular components responsible for its therapeutic benefit have been explored. Stem cells, as well as most cells, release extracellular vesicles such as exosomes, which are nanovesicles able to target specific cell types and thus to modify their function by delivering proteins, lipids, and nucleic acids. Exosomes have recently been tested in vivo and in vitro as therapeutic conveyors for the treatment of diseases. As such, they could be engineered to target specific populations of cells within the CNS. Considering the fact that many degenerative brain diseases have an impact on adult neurogenesis, we discuss how the modulation of the adult neurogenic niches may be a therapeutic target of stem cell-derived exosomes. These novel approaches should be examined in cellular and animal models to provide better, more effective, and specific therapeutic tools in the future. PMID:27195011

  10. HPV-E7 Delivered by Engineered Exosomes Elicits a Protective CD8+ T Cell-Mediated Immune Response

    Directory of Open Access Journals (Sweden)

    Paola Di Bonito

    2015-03-01

    Full Text Available We developed an innovative strategy to induce a cytotoxic T cell (CTL immune response against protein antigens of choice. It relies on the production of exosomes, i.e., nanovesicles spontaneously released by all cell types. We engineered the upload of huge amounts of protein antigens upon fusion with an anchoring protein (i.e., HIV-1 Nefmut, which is an inactive protein incorporating in exosomes at high levels also when fused with foreign proteins. We compared the immunogenicity of engineered exosomes uploading human papillomavirus (HPV-E7 with that of lentiviral virus-like particles (VLPs incorporating equivalent amounts of the same antigen. These exosomes, whose limiting membrane was decorated with VSV-G, i.e., an envelope protein inducing pH-dependent endosomal fusion, proved to be as immunogenic as the cognate VLPs. It is noteworthy that the immunogenicity of the engineered exosomes remained unaltered in the absence of VSV-G. Most important, we provide evidence that the inoculation in mouse of exosomes uploading HPV-E7 induces production of anti-HPV E7 CTLs, blocks the growth of syngeneic tumor cells inoculated after immunization, and controls the development of tumor cells inoculated before the exosome challenge. These results represent the proof-of-concept about both feasibility and efficacy of the Nefmut-based exosome platform for the induction of CD8+ T cell immunity.

  11. Development of exosome surface display technology in living human cells.

    Science.gov (United States)

    Stickney, Zachary; Losacco, Joseph; McDevitt, Sophie; Zhang, Zhiwen; Lu, Biao

    2016-03-25

    Surface display technology is an emerging key player in presenting functional proteins for targeted drug delivery and therapy. Although a number of technologies exist, a desirable mammalian surface display system is lacking. Exosomes are extracellular vesicles that facilitate cell-cell communication and can be engineered as nano-shuttles for cell-specific delivery. In this study, we report the development of a novel exosome surface display technology by exploiting mammalian cell secreted nano-vesicles and their trans-membrane protein tetraspanins. By constructing a set of fluorescent reporters for both the inner and outer surface display on exosomes at two selected sites of tetraspanins, we demonstrated the successful exosomal display via gene transfection and monitoring fluorescence in vivo. We subsequently validated our system by demonstrating the expected intracellular partitioning of reporter protein into sub-cellular compartments and secretion of exosomes from human HEK293 cells. Lastly, we established the stable engineered cells to harness the ability of this robust system for continuous production, secretion, and uptake of displayed exosomes with minimal impact on human cell biology. In sum, our work paved the way for potential applications of exosome, including exosome tracking and imaging, targeted drug delivery, as well as exosome-mediated vaccine and therapy.

  12. Exosomes as a tumor immune escape mechanism: possible therapeutic implications

    Directory of Open Access Journals (Sweden)

    Hanley Harold H

    2008-07-01

    Full Text Available Abstract Advances in cancer therapy have been substantial in terms of molecular understanding of disease mechanisms, however these advances have not translated into increased survival in the majority of cancer types. One unsolved problem in current cancer therapeutics is the substantial immune suppression seen in patients. Conventionally, investigations in this area have focused on antigen-nonspecific immune suppressive molecules such as cytokines and T cell apoptosis inducing molecules such as Fas ligand. More recently, studies have demonstrated nanovesicle particles termed exosomes are involved not only in stimulation but also inhibition of immunity in physiological conditions. Interestingly, exosomes secreted by cancer cells have been demonstrated to express tumor antigens, as well as immune suppressive molecules such as PD-1L and FasL. Concentrations of exosomes from plasma of cancer patients have been associated with spontaneous T cell apoptosis, which is associated in some situations with shortened survival. In this paper we place the "exosome-immune suppression" concept in perspective of other tumor immune evasion mechanisms. We conclude by discussing a novel therapeutic approach to cancer immune suppression by extracorporeal removal of exosomes using hollow fiber filtration technology

  13. Tumor Microenvironment Modulation via Gold Nanoparticles Targeting Malicious Exosomes: Implications for Cancer Diagnostics and Therapy

    Directory of Open Access Journals (Sweden)

    Catarina Roma-Rodrigues

    2017-01-01

    Full Text Available Exosomes are nanovesicles formed in the endosomal pathway with an important role in paracrine and autocrine cell communication. Exosomes secreted by cancer cells, malicious exosomes, have important roles in tumor microenvironment maturation and cancer progression. The knowledge of the role of exosomes in tumorigenesis prompted a new era in cancer diagnostics and therapy, taking advantage of the use of circulating exosomes as tumor biomarkers due to their stability in body fluids and targeting malignant exosomes’ release and/or uptake to inhibit or delay tumor development. In recent years, nanotechnology has paved the way for the development of a plethora of new diagnostic and therapeutic platforms, fostering theranostics. The unique physical and chemical properties of gold nanoparticles (AuNPs make them suitable vehicles to pursuit this goal. AuNPs’ properties such as ease of synthesis with the desired shape and size, high surface:volume ratio, and the possibility of engineering their surface as desired, potentiate AuNPs’ role in nanotheranostics, allowing the use of the same formulation for exosome detection and restraining the effect of malicious exosomes in cancer progression.

  14. Maximizing exosome colloidal stability following electroporation.

    Science.gov (United States)

    Hood, Joshua L; Scott, Michael J; Wickline, Samuel A

    2014-03-01

    Development of exosome-based semisynthetic nanovesicles for diagnostic and therapeutic purposes requires novel approaches to load exosomes with cargo. Electroporation has previously been used to load exosomes with RNA. However, investigations into exosome colloidal stability following electroporation have not been considered. Herein, we report the development of a unique trehalose pulse media (TPM) that minimizes exosome aggregation following electroporation. Dynamic light scattering (DLS) and RNA absorbance were employed to determine the extent of exosome aggregation and electroextraction post electroporation in TPM compared to common PBS pulse media or sucrose pulse media (SPM). Use of TPM to disaggregate melanoma exosomes post electroporation was dependent on both exosome concentration and electric field strength. TPM maximized exosome dispersal post electroporation for both homogenous B16 melanoma and heterogeneous human serum-derived populations of exosomes. Moreover, TPM enabled heavy cargo loading of melanoma exosomes with 5nm superparamagnetic iron oxide nanoparticles (SPION5) while maintaining original exosome size and minimizing exosome aggregation as evidenced by transmission electron microscopy. Loading exosomes with SPION5 increased exosome density on sucrose gradients. This provides a simple, label-free means of enriching exogenously modified exosomes and introduces the potential for MRI-driven theranostic exosome investigations in vivo.

  15. Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma.

    Science.gov (United States)

    Kalra, Hina; Adda, Christopher G; Liem, Michael; Ang, Ching-Seng; Mechler, Adam; Simpson, Richard J; Hulett, Mark D; Mathivanan, Suresh

    2013-11-01

    Exosomes are nanovesicles released by a variety of cells and are detected in body fluids including blood. Recent studies have highlighted the critical application of exosomes as personalized targeted drug delivery vehicles and as reservoirs of disease biomarkers. While these research applications have created significant interest and can be translated into practice, the stability of exosomes needs to be assessed and exosome isolation protocols from blood plasma need to be optimized. To optimize methods to isolate exosomes from blood plasma, we performed a comparative evaluation of three exosome isolation techniques (differential centrifugation coupled with ultracentrifugation, epithelial cell adhesion molecule immunoaffinity pull-down, and OptiPrep(TM) density gradient separation) using normal human plasma. Based on MS, Western blotting and microscopy results, we found that the OptiPrep(TM) density gradient method was superior in isolating pure exosomal populations, devoid of highly abundant plasma proteins. In addition, we assessed the stability of exosomes in plasma over 90 days under various storage conditions. Western blotting analysis using the exosomal marker, TSG101, revealed that exosomes are stable for 90 days. Interestingly, in the context of cellular uptake, the isolated exosomes were able to fuse with target cells revealing that they were indeed biologically active.

  16. Exosome-mediated delivery of functionally active miRNA-155 inhibitor to macrophages.

    Science.gov (United States)

    Momen-Heravi, Fatemeh; Bala, Shashi; Bukong, Terence; Szabo, Gyongyi

    2014-10-01

    Exosomes, membranous nanovesicles, naturally carry bio-macromolecules and play pivotal roles in both physiological intercellular crosstalk and disease pathogenesis. Here, we showed that B cell-derived exosomes can function as vehicles to deliver exogenous miRNA-155 mimic or inhibitor into hepatocytes or macrophages, respectively. Stimulation of B cells significantly increased exosome production. Unlike in parental cells, baseline level of miRNA-155 was very low in exosomes derived from stimulated B cells. Exosomes loaded with a miRNA-155 mimic significantly increased miRNA-155 levels in primary mouse hepatocytes and the liver of miRNA-155 knockout mice. Treatment of RAW macrophages with miRNA-155 inhibitor loaded exosomes resulted in statistically significant reduction in LPS-induced TNFα production and partially prevented LPS-induced decrease in SOCS1 mRNA levels. Furthermore, exosome-mediated miRNA-155 inhibitor delivery resulted in functionally more efficient inhibition and less cellular toxicity compared to conventional transfection methods. Similar approaches could be useful in modification of target biomolecules in vitro and in vivo. From the clinical editor: In this study, exosome-based delivery of miRNA-155 mimicker or inhibitor was found to have significant biological response in hepatocytes and macrophages. Exosome-based approaches may be useful in the modification of other target biomolecules.

  17. Exosomes: Implications in HIV-1 Pathogenesis.

    Science.gov (United States)

    Madison, Marisa N; Okeoma, Chioma M

    2015-07-20

    Exosomes are membranous nanovesicles of endocytic origin that carry host and pathogen derived genomic, proteomic, and lipid cargos. Exosomes are secreted by most cell types into the extracellular milieu and are subsequently internalized by recipient cells. Upon internalization, exosomes condition recipient cells by donating their cargos and/or activating various signal transduction pathways, consequently regulating physiological and pathophysiological processes. The role of exosomes in viral pathogenesis, especially human immunodeficiency virus type 1 [HIV-1] is beginning to unravel. Recent research reports suggest that exosomes from various sources play important but different roles in the pathogenesis of HIV-1. From these reports, it appears that the source of exosomes is the defining factor for the exosomal effect on HIV-1. In this review, we will describe how HIV-1 infection is modulated by exosomes and in turn how exosomes are targeted by HIV-1 factors. Finally, we will discuss potentially emerging therapeutic options based on exosomal cargos that may have promise in preventing HIV-1 transmission.

  18. Exosomes: Implications in HIV-1 Pathogenesis

    Directory of Open Access Journals (Sweden)

    Marisa N. Madison

    2015-07-01

    Full Text Available Exosomes are membranous nanovesicles of endocytic origin that carry host and pathogen derived genomic, proteomic, and lipid cargos. Exosomes are secreted by most cell types into the extracellular milieu and are subsequently internalized by recipient cells. Upon internalization, exosomes condition recipient cells by donating their cargos and/or activating various signal transduction pathways, consequently regulating physiological and pathophysiological processes. The role of exosomes in viral pathogenesis, especially human immunodeficiency virus type 1 [HIV-1] is beginning to unravel. Recent research reports suggest that exosomes from various sources play important but different roles in the pathogenesis of HIV-1. From these reports, it appears that the source of exosomes is the defining factor for the exosomal effect on HIV-1. In this review, we will describe how HIV-1 infection is modulated by exosomes and in turn how exosomes are targeted by HIV-1 factors. Finally, we will discuss potentially emerging therapeutic options based on exosomal cargos that may have promise in preventing HIV-1 transmission.

  19. Exosomes as new vesicular lipid transporters involved in cell-cell communication and various pathophysiologies.

    Science.gov (United States)

    Record, Michel; Carayon, Kevin; Poirot, Marc; Silvente-Poirot, Sandrine

    2014-01-01

    Exosomes are nanovesicles that have emerged as a new intercellular communication system between an intracellular compartment of a donor cell towards the periphery or an internal compartment of a recipient cell. The bioactivity of exosomes resides not only in their protein and RNA contents but also in their lipidic molecules. Exosomes display original lipids organized in a bilayer membrane and along with the lipid carriers such as fatty acid binding proteins that they contain, exosomes transport bioactive lipids. Exosomes can vectorize lipids such as eicosanoids, fatty acids, and cholesterol, and their lipid composition can be modified by in-vitro manipulation. They also contain lipid related enzymes so that they can constitute an autonomous unit of production of various bioactive lipids. Exosomes can circulate between proximal or distal cells and their fate can be regulated in part by lipidic molecules. Compared to their parental cells, exosomes are enriched in cholesterol and sphingomyelin and their accumulation in cells might modulate recipient cell homeostasis. Exosome release from cells appears to be a general biological process. They have been reported in all biological fluids from which they can be recovered and can be monitors of specific pathophysiological situations. Thus, the lipid content of circulating exosomes could be useful biomarkers of lipid related diseases. Since the first lipid analysis of exosomes ten years ago detailed knowledge of exosomal lipids has accumulated. The role of lipids in exosome fate and bioactivity and how they constitute an additional lipid transport system are considered in this review.

  20. Biomimetic aqueous-core lipid nanoballoons integrating a multiple emulsion formulation: a suitable housing system for viable lytic bacteriophages.

    Science.gov (United States)

    Balcão, Victor M; Glasser, Cássia A; Chaud, Marco V; del Fiol, Fernando S; Tubino, Matthieu; Vila, Marta M D C

    2014-11-01

    The emergence of antibiotic-resistant bacterial strains and the weak penetration of antibiotics into bacterial biofilms put an emphasis in the need for safe and effective alternatives for antimicrobial treatments. The application of strictly lytic bacteriophages (or phages) has been proposed as an alternative (or complement) to conventional antibiotics, allowing release of the natural predators of bacteria directly to the site of infection. In the present research effort, production of bacteriophage derivatives (starting from lytic phage particle isolates), encompassing full stabilization of their three-dimensional structure, has been attempted via housing said bacteriophage particles within lipid nanovesicles integrating a multiple water-in-oil-in-water (W/O/W) emulsion. As a proof-of-concept for the aforementioned strategy, bacteriophage particles with broad lytic spectrum were entrapped within the aqueous core of lipid nanoballoons integrating a W/O/W multiple emulsion. Long-term storage of the multiple emulsions produced did not lead to leaching of phage particles, thus proving the effectiveness of the encapsulation procedure.

  1. Embryonic–maternal cross-talk via exosomes: potential implications

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    Saadeldin IM

    2015-07-01

    Full Text Available Islam M Saadeldin,1 Hyun Ju Oh,2 Byeong Chun Lee2,3 1Department of Physiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt; 2Department of Theriogenology and Biotechnology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul, Republic of Korea; 3Institute of Green Bio Science and Technology, Seoul National University, Pyeong Chang, Kangwon do, Republic of KoreaAbstract: A myriad of locally produced factors into the microenvironment of the reproductive tract is regulated, not one-way but rather, through embryonic–maternal cross-talk. In this minireview, we focused on the exosomes, which are cell-derived vesicles of 30–100 nm in diameter, as a communicating language facilitating this dialog. These nanovesicles are secreted from preimplantation embryos, oviduct epithelium, and endometrium as well as from the placenta, and contain proteins, messenger RNA (mRNA, microRNA, and DNA cargoes, and have pleiotropic effects on both embryonic and maternal environments. A better understanding of the molecular mechanisms mediating this cross-talk will lead to the development of new regulating agents, with novel diagnostic, biological, and therapeutic potential for either supporting or hindering the normal reproductive functions. Keywords: embryo, endometrium, placenta, mRNA, miRNA

  2. Neutrophil-Derived Exosomes: A New Mechanism Contributing to Airway Smooth Muscle Remodeling.

    Science.gov (United States)

    Vargas, Amandine; Roux-Dalvai, Florence; Droit, Arnaud; Lavoie, Jean-Pierre

    2016-09-01

    Neutrophils infiltrate the airways of patients with asthma of all severities, yet their role in the pathogenesis of asthma and their contribution to airway remodeling is largely unknown. We hypothesized that neutrophils modulate airway smooth muscle (ASM) proliferation in asthma by releasing bioactive exosomes. These newly discovered nano-sized vesicles have the capacity to modulate immune responses, cell migration, cell differentiation, and other aspects of cell-to-cell communication. The aim of the study is to determine whether bioactive exosomes are released by neutrophils, and, if so, characterize their proteomic profile and evaluate their capacity to modulate ASM cell proliferation. Exosomes were isolated from equine neutrophil supernatants by differential centrifugation and filtration methods, followed by size-exclusion chromatography. Nanovesicles were characterized using electron microscopy, particle size determination, and proteomic analyses. Exosomes were cocultured with ASM cells and analyzed for exosome internalization by confocal microscopy. ASM proliferation was measured using an impedance-based system. Neutrophils release exosomes that have characteristic size, morphology, and exosomal markers. We identified 271 proteins in exosomes from both LPS and unstimulated neutrophils, and 16 proteins that were differentially expressed, which carried proteins associated with immune response and positive regulation of cell communication. Furthermore, neutrophil-derived exosomes were rapidly internalized by ASM cells and altered their proliferative properties. Upon stimulation of LPS, neutrophil-derived exosomes can enhance the proliferation of ASM cells and could therefore play an important role in the progression of asthma and promoting airway remodeling in severe and corticosteroid-insensitive patients with asthma.

  3. Role of exosomes released by dendritic cells and/or by tumor targets: Regulation of NK cell plasticity.

    Directory of Open Access Journals (Sweden)

    Katrin S. Reiners

    2014-03-01

    Full Text Available Exosomes are endosomal-derived nanovesicles released by normal and tumor cells, which transfer functionally active proteins, lipids and nucleic acids between cells. They are important mediators of intercellular communication and act on the adjacent stroma as well as in the periphery. Recently, exosomes have been recognized to play a pathophysiological role in various diseases such as cancer or infectious diseases. Tumor cell-derived exosomes (Tex have been shown to act as tumor promotors by educating non-malignant cells to provide a tumor supporting microenvironment, which helps to circumvent immune detection by the host and supports metastasis. However, Tex with anti-tumor, immune-activating properties were also described reflecting the complexity of exosomes.Here, we assess the role of extracellular microvesicles/exosomes as messengers affecting NK cell function in health and disease and discuss the molecular basis for the differential impact of exosomes on NK cell activity. The molecular composition/load of exosomes and the mechanisms regulating their release remain unclear and need to be further analyzed to facilitate the development of new treatment options targeting the exosomal machinery.

  4. Digital Detection of Exosomes by Interferometric Imaging

    Science.gov (United States)

    Daaboul, George G.; Gagni, Paola; Benussi, Luisa; Bettotti, Paolo; Ciani, Miriam; Cretich, Marina; Freedman, David S.; Ghidoni, Roberta; Ozkumur, Ayca Yalcin; Piotto, Chiara; Prosperi, Davide; Santini, Benedetta; Ünlü, M. Selim; Chiari, Marcella

    2016-01-01

    Exosomes, which are membranous nanovesicles, are actively released by cells and have been attributed to roles in cell-cell communication, cancer metastasis, and early disease diagnostics. The small size (30–100 nm) along with low refractive index contrast of exosomes makes direct characterization and phenotypical classification very difficult. In this work we present a method based on Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) that allows multiplexed phenotyping and digital counting of various populations of individual exosomes (>50 nm) captured on a microarray-based solid phase chip. We demonstrate these characterization concepts using purified exosomes from a HEK 293 cell culture. As a demonstration of clinical utility, we characterize exosomes directly from human cerebrospinal fluid (hCSF). Our interferometric imaging method could capture, from a very small hCSF volume (20 uL), nanoparticles that have a size compatible with exosomes, using antibodies directed against tetraspanins. With this unprecedented capability, we foresee revolutionary implications in the clinical field with improvements in diagnosis and stratification of patients affected by different disorders. PMID:27853258

  5. Stable J-aggregation enabled dual photoacoustic and fluorescence nanoparticles for intraoperative cancer imaging

    Science.gov (United States)

    Shakiba, Mojdeh; Ng, Kenneth K.; Huynh, Elizabeth; Chan, Harley; Charron, Danielle M.; Chen, Juan; Muhanna, Nidal; Foster, F. Stuart; Wilson, Brian C.; Zheng, Gang

    2016-06-01

    J-aggregates display nanoscale optical properties which enable their use in fluorescence and photoacoustic imaging applications. However, control over their optical properties in an in vivo setting is hampered by the conformational lability of the J-aggregate structure in complex biological environments. J-aggregating nanoparticles (JNP) formed by self-assembly of bacteriopheophorbide-lipid (Bchl-lipid) in lipid nanovesicles represents a novel strategy to stabilize J-aggregates for in vivo bioimaging applications. We find that 15 mol% Bchl-lipid embedded within a saturated phospholipid bilayer vesicle was optimal in terms of maximizing Bchl-lipid dye loading, while maintaining a spherical nanoparticle morphology and retaining spectral properties characteristic of J-aggregates. The addition of cholesterol maintains the stability of the J-aggregate absorption band for up to 6 hours in the presence of 90% FBS. In a proof-of-concept experiment, we successfully applied JNPs as a fluorescence contrast agent for real-time intraoperative detection of metastatic lymph nodes in a rabbit head-and-neck cancer model. Lymph node metastasis delineation was further verified by visualizing the JNP within the excised lymph node using photoacoustic imaging. Using JNPs, we demonstrate the possibility of using J-aggregates as fluorescence and photoacoustic contrast agents and may potentially spur the development of other nanomaterials that can stably induce J-aggregation for in vivo cancer bioimaging applications.J-aggregates display nanoscale optical properties which enable their use in fluorescence and photoacoustic imaging applications. However, control over their optical properties in an in vivo setting is hampered by the conformational lability of the J-aggregate structure in complex biological environments. J-aggregating nanoparticles (JNP) formed by self-assembly of bacteriopheophorbide-lipid (Bchl-lipid) in lipid nanovesicles represents a novel strategy to stabilize J

  6. Dromedary milk exosomes as mammary transcriptome nano-vehicle: Their isolation, vesicular and phospholipidomic characterizatio

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    Aya M. Yassin

    2016-09-01

    Full Text Available Exosomes are extracellular nanovesicles that play a role in cellular trafficking and communication. Camel milk exosomes might carry the potential of recovery of several illnesses that coins the dromedary milk. This study shows for the first time their isolation and fine characterization. The differential ultracentrifugation was used for their isolation. Their recovery from dromedary milk during different lactation periods was evaluated. The vesicular characterization and stability testing of the recovered exosome were examined by transmission electron microscopy (TEM. The proteome footprinting was resolved by gel electrophoresis prior to their specific protein biomarker analysis. The immunoblotting of their specific protein biomarker TSG101 unexpectedly revealed a truncated 35 KDa protein specific for dromedary milk exosome rather than the previously reported 43 KDa mammalian one. The reversed-phase HPLC screening of their phospholipid makeup was compared with that of cattle milk exosomes at different lactation periods. Since dromedary milk exosomes reflect their mammary transcriptome outcome, further assessment of their content of αs1casein, αs2casein β-casein κ-casein mRNAs parallel with a constitutive glyceraldehyde dehydrogenase (GAPD gene was performed using real-time PCR. The TEM scanning indicated that dromedary milk exosomes are freeze-stress unstable homogeneous with average size of 30 nm. There was no significant difference in expression level of different casein genes in mid lactation period in dromedary milk exosomes over late lactation period. The phospholipidomic survey proved that phosphatidylcholine is the major candidate of the examined phospholipids in dromedary milk exosomes. The obtained data give novel interpretation about the content of camel milk exosomes with possible insight for use as potentially-safe nano carrier.

  7. A strategy of antigen incorporation into exosomes: comparing cross-presentation levels of antigens delivered by engineered exosomes and by lentiviral virus-like particles.

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    Lattanzi, Laura; Federico, Maurizio

    2012-11-26

    Among strategies aimed at developing new nanoparticle-based vaccines, exosomes hold much promise. They are nanovesicles released by basically all eukaryotic cell types originating from intraluminal vesicles which accumulate in multivesicular bodies. Exosomes have immunogenic properties whose strength correlates with the amounts of associated antigens. Engineering antigens to target them in exosomes represents the last frontier in terms of nanoparticle-based vaccines. Here we report a new method to incorporate protein antigens in exosomes relying on the unique properties of a mutant of the HIV-1 Nef protein, Nef(mut). This is a biologically inactive mutant we found incorporating into exosomes at high levels also when fused at its C-terminus with foreign proteins. We compared both biochemical and antigenic properties of Nef(mut) exosomes with those of previously characterized Nef(mut) -based lentiviral virus-like particles (VLPs). We found that exosomes incorporate Nef(mut) and fusion protein derivatives with similar efficiency of VLPs. When an envelope fusion protein was associated with both exosomes and VLPs to favor cross-presentation of associated antigens, Nef(mut) and its derivatives incorporated in exosomes were cross-presented at levels at least similar to what observed when the antigens were delivered by engineered VLPs. This occurred despite exosomes entered target cells with an apparent lower efficiency than VLPs. The unique properties of HIV-1 Nef(mut) in terms of exosome incorporation efficiency, carrier of foreign antigens, and lack of anti-cellular effects open the way toward the development of a flexible, safe, cost-effective exosome-based CD8(+) T cell vaccine platform.

  8. Development of curcumin loaded sodium hyaluronate immobilized vesicles (hyalurosomes) and their potential on skin inflammation and wound restoring.

    Science.gov (United States)

    Manca, M L; Castangia, I; Zaru, M; Nácher, A; Valenti, D; Fernàndez-Busquets, X; Fadda, A M; Manconi, M

    2015-12-01

    In the present work new highly biocompatible nanovesicles were developed using polyanion sodium hyaluronate to form polymer immobilized vesicles, so called hyalurosomes. Curcumin, at high concentration was loaded into hyalurosomes and physico-chemical properties and in vitro/in vivo performances of the formulations were compared to those of liposomes having the same lipid and drug content. Vesicles were prepared by direct addition of dispersion containing the polysaccharide sodium hyaluronate and the polyphenol curcumin to a commercial mixture of soy phospholipids, thus avoiding the use of organic solvents. An extensive study was carried out on the physico-chemical features and properties of curcumin-loaded hyalurosomes and liposomes. Cryogenic transmission electron microscopy and small-angle X-ray scattering showed that vesicles were spherical, uni- or oligolamellar and small in size (112-220 nm). The in vitro percutaneous curcumin delivery studies on intact skin showed an improved ability of hyalurosomes to favour a fast drug deposition in the whole skin. Hyalurosomes as well as liposomes were biocompatible, protected in vitro human keratinocytes from oxidative stress damages and promoted tissue remodelling through cellular proliferation and migration. Moreover, in vivo tests underlined a good effectiveness of curcumin-loaded hyalurosomes to counteract 12-O-tetradecanoilphorbol (TPA)-produced inflammation and injuries, diminishing oedema formation, myeloperoxydase activity and providing an extensive skin reepithelization. Thanks to the one-step and environmentally-friendly preparation method, component biocompatibility and safety, good in vitro and in vivo performances, the hyalurosomes appear as promising nanocarriers for cosmetic and pharmaceutical applications.

  9. Vibrational spectroscopic studies of newly developed synthetic biopolymers.

    Science.gov (United States)

    Bista, Rajan K; Bruch, Reinhard F; Covington, Aaron M

    2010-05-01

    Vibrational spectroscopic techniques such as near-infrared (NIR), Fourier transform infrared (FTIR), and Raman spectroscopy are valuable diagnostic tools that can be used to elucidate comprehensive structural information of numerous biological samples. In this review article, we have highlighted the advantages of nanotechnology and biophotonics in conjunction with vibrational spectroscopic techniques in order to understand the various aspects of new kind of synthetic biopolymers termed as polyethylene glycol (PEG)ylated lipids. In contrast to conventional phospholipids, these novel lipids spontaneously form liposomes or nanovesicles upon hydration, without the supply of external activation energy. The amphiphiles considered in this study differ in their hydrophobic acyl chain length and contain different units of PEG hydrophilic headgroups. We have further explored the thermotropic phase behaviors and associated changes in the conformational order/disorder of such lipids by using variable-temperature FTIR and Raman spectroscopy. Phase transition temperature profiles and correlation between various spectral indicators have been identified by either monitoring the shifts in the vibrational peak positions or plotting vibrational peak intensity ratios in the C--H stretching region as a function of temperature. To supplement our observations of phase transformations, a thermodynamic approach known as differential scanning calorimetry (DSC) has been applied and revealed a good agreement with the infrared and Raman spectroscopic data. Finally, the investigation of thermal properties of lipids is extremely crucial for numerous purposes, thus the results obtained in this work may find application in a wide variety of studies including the development of PEGylated lipid based drug and substances delivery vehicles.

  10. Proteomic analysis of exosomes from nasopharyngeal carcinoma cell identifies intercellular transfer of angiogenic proteins

    KAUST Repository

    Chan, Yuk-kit

    2015-04-01

    Exosomes, a group of secreted extracellular nanovesicles containing genetic materials and signaling molecules, play a critical role in intercellular communication. During tumorigenesis, exosomes have been demonstrated to promote tumor angiogenesis and metastasis while their biological functions in nasopharyngeal carcinoma (NPC) are poorly understood. In this study, we focused on the role of NPC-derived exosomes on angiogenesis. Exosomes derived from the NPC C666-1 cells and immortalized nasopharyngeal epithelial cells (NP69 and NP460) were isolated using ultracentrifugation. The molecular profile and biophysical characteristics of exosomes were verified by Western blotting, sucrose density gradient, and electron microscopy. We showed that the C666-1 exosomes (10 and 20 μg/ml) could significantly increase the tubulogenesis, migration and invasion of human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner. Subsequently, an iTRAQ-based quantitative proteomics was used to identify the differentially expressed proteins in C666-1 exosomes. Among the 640 identified proteins, 51 and 89 proteins were considered as up- and down-regulated (≥ 1.5-fold variations) in C666-1 exosomes compared to the normal counterparts, respectively. As expected, pro-angiogenic proteins including intercellular adhesion molecule-1 (ICAM-1) and CD44 variant isoform 5 (CD44v5) are among the up-regulated proteins, whereas angio-suppressive protein, thrombospondin-1 (TSP-1) was down-regulated in C666-1 exosomes. Further confocal microscopic study and Western blotting clearly demonstrated that the alteration of ICAM-1, and TSP-1 expressions in recipient HUVECs are due to internalization of exosomes. Taken together, these data strongly indicated the critical roles of identified angiogenic proteins in the involvement of exosomes-induced angiogenesis, which could potentially be developed as therapeutic targets in future. This article is protected by copyright. All rights reserved.

  11. Emerging roles of exosomes in normal and pathological conditions. New insights for diagnosis and therapeutic applications

    Directory of Open Access Journals (Sweden)

    Julieta eDe Toro

    2015-05-01

    Full Text Available From the time when they were first described in the 1970s by the group of Johnstone and Stahl, exosomes are a target of constant research. Exosomes belong to the family of nano-vesicles which are of great interest for their many functions and potential for diagnosis and therapy in multiples diseases. Exosomes originate from the intraluminal vesicles of late endosomal compartments named multivesicular bodies and the fusion of these late endosomes with the cell membrane result in the release of the vesicles into the extracellular compartment. Moreover, their generation can be induced by many factors including extracellular stimuli, such as microbial attack and other stress conditions. The primary role attributed to exosomes was the removal of unnecessary proteins from the cells. Now, several studies have demonstrated that exosomes are involved in cell-cell communication, even though their biological function is not completely clear.The participation of exosomes in cancer is the field of microvesicle research that has expanded more over the last years. Evidence proving that exosomes derived from tumor-pulsed dendritic cells, neoplastic cells and malignant effusions, are able to present antigens to T‐cells, has led to numerous studies using them as cell free cancer vaccines.Since exosomes derive from all cell types, they contain proteins, lipids and miRNA capable of regulating a variety of target genes. Much research is being conducted, which focuses on the employment of these vesicles as biomarkers in the diagnosis of cancer in addition to innovative biomarkers for diagnosis, prognosis and management of cardiovascular diseases. Interesting findings indicating the role of exosomes in the pathogenesis of several diseases have encouraged researchers to consider their therapeutic potential not only in oncology but also in the treatment of autoimmune syndromes and neurodegenerative disorders such as Alzheimer´s and Parkinson´s disease; in addition

  12. Development of a rapid lateral flow immunoassay test for detection of exosomes previously enriched from cell culture medium and body fluids

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    Myriam Oliveira-Rodríguez

    2016-08-01

    Full Text Available Exosomes are cell-secreted nanovesicles (40–200 nm that represent a rich source of novel biomarkers in the diagnosis and prognosis of certain diseases. Despite the increasingly recognized relevance of these vesicles as biomarkers, their detection has been limited due in part to current technical challenges in the rapid isolation and analysis of exosomes. The complexity of the development of analytical platforms relies on the heterogeneous composition of the exosome membrane. One of the most attractive tests is the inmunochromatographic strips, which allow rapid detection by unskilled operators. We have successfully developed a novel lateral flow immunoassay (LFIA for the detection of exosomes based on the use of tetraspanins as targets. We have applied this platform for the detection of exosomes purified from different sources: cell culture supernatants, human plasma and urine. As proof of concept, we explored the analytical potential of this LFIA platform to accurately quantify exosomes purified from a human metastatic melanoma cell line. The one-step assay can be completed in 15 min, with a limit of detection of 8.54×105 exosomes/µL when a blend of anti-CD9 and anti-CD81 were selected as capture antibodies and anti-CD63 labelled with gold nanoparticles as detection antibody. Based on our results, this platform could be well suited to be used as a rapid exosome quantification tool, with promising diagnostic applications, bearing in mind that the detection of exosomes from different sources may require adaptation of the analytical settings to their specific composition.

  13. Supramolecular nanoparticles generated by the self-assembly of polyrotaxanes for antitumor drug delivery

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    Liu R

    2012-10-01

    Full Text Available Rong Liu,1,2,* Yusi Lai,1,* Bin He,1 Yuan Li,1 Gang Wang,1 Shuang Chang,1 Zhongwei Gu1 1National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, China; 2Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China*These authors contributed equally to this paperAbstract: A new approach of fabricating supramolecular nanoparticles generated by self-assembly polyrotaxanes for antitumor drug delivery has been reported. Cinnamic-acid-modified poly(ethylene glycol chains were threaded in a-cyclodextrins to form polyrotaxanes. The polyrotaxanes self-assembled supramolecular nanoparticles. The morphology of the nanoparticles was changed from nanovesicle to micelle after the antitumor drug, doxorubicin, was loaded. The release profile of the drug-loaded nanoparticles was investigated, and it was found that the sustaining release time could last for 32 hours. The drug-loaded nanoparticles were co-cultured with mouse 4T1 breast cancer cells with a drug concentration of 10 µg/mL; the cell survival rate was 3.3% after a 72-hour incubation. In an in vivo study of breast cancer in a mouse model, the drug-loaded nanoparticles were injected in the tail veins of mice with a dose of 5 mg/kg body weight. The tumor inhibition rate of drug-loaded nanoparticles was 53%, which was better than that of doxorubicin hydrochloride. The cardiac toxicity of doxorubicin was decreased greatly after the encapsulation into supramolecular polyrotaxane nanoparticles.Keywords: polyrotaxane, self-assembly, nanoparticle, doxorubicin, supermolecular

  14. On-chip immunoelectrophoresis of extracellular vesicles released from human breast cancer cells.

    Science.gov (United States)

    Akagi, Takanori; Kato, Kei; Kobayashi, Masashi; Kosaka, Nobuyoshi; Ochiya, Takahiro; Ichiki, Takanori

    2015-01-01

    Extracellular vesicles (EVs) including exosomes and microvesicles have attracted considerable attention in the fields of cell biology and medicine. For a better understanding of EVs and further exploration of their applications, the development of analytical methods for biological nanovesicles has been required. In particular, considering the heterogeneity of EVs, methods capable of measuring individual vesicles are desired. Here, we report that on-chip immunoelectrophoresis can provide a useful method for the differential protein expression profiling of individual EVs. Electrophoresis experiments were performed on EVs collected from the culture supernatant of MDA-MB-231 human breast cancer cells using a measurement platform comprising a microcapillary electrophoresis chip and a laser dark-field microimaging system. The zeta potential distribution of EVs that reacted with an anti-human CD63 (exosome and microvesicle marker) antibody showed a marked positive shift as compared with that for the normal immunoglobulin G (IgG) isotype control. Thus, on-chip immunoelectrophoresis could sensitively detect the over-expression of CD63 glycoproteins on EVs. Moreover, to explore the applicability of on-chip immunoelectrophoresis to cancer diagnosis, EVs collected from the blood of a mouse tumor model were analyzed by this method. By comparing the zeta potential distributions of EVs after their immunochemical reaction with normal IgG, and the anti-human CD63 and anti-human CD44 (cancer stem cell marker) antibodies, EVs of tumor origin circulating in blood were differentially detected in the real sample. The result indicates that the present method is potentially applicable to liquid biopsy, a promising approach to the low-invasive diagnosis of cancer.

  15. An emerging class of amphiphilic dendrimers for pharmaceutical and biomedical applications: Janus amphiphilic dendrimers.

    Science.gov (United States)

    Sikwal, Dhiraj R; Kalhapure, Rahul S; Govender, Thirumala

    2017-01-15

    In recent years, a new class of dendrimer, known as Janus dendrimers (JDs), has attracted much attention due to their different structures and properties to the conventional symmetric forms. The broken symmetry of JDs offers the opportunity to form complex self-assembled materials, and presents new sets of properties that are presently inconceivable for homogeneous or symmetrical dendrimers. Due to their unique features, JDs have a promising future in pharmaceutical and biomedical fields, as seen from the recent interest in their application in conjugating multiple drugs and targeting moieties, forming supramolecular hydrogels, enabling micellar delivery systems, and preparing nano-vesicles, known as dendrimersomes, for drug encapsulation. The present paper is the first review, with an emphasis on various emerging applications of JDs, in the drug delivery and biomedical field reported so far. In addition, the paper describes different synthetic methods for producing JDs that can guide the design of new biocompatible forms with pharmacological activities, and that have the potential to be nano drug delivery vehicles. Furthermore, future studies to optimize the applications of JDs in drug delivery sciences and biomedical field to realize their potential to treat various disease conditions are identified and highlighted. Overall, this review identifies the current status of JDs in terms of their synthesis and applications, as well as the future research for their translation into macromolecules for clinical applications to solve health problems. It highlights the future combined efforts needed to be taken by dendrimer chemists, formulation scientist and microbiologists to develop novel antibacterials and nanomedicines from JDs. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Liposome and niosome preparation using a membrane contactor for scale-up.

    Science.gov (United States)

    Pham, Thi Thuy; Jaafar-Maalej, Chiraz; Charcosset, Catherine; Fessi, Hatem

    2012-06-01

    The scaling-up ability of liposome and niosome production, from laboratory scale using a syringe-pump device to a pilot scale using the membrane contactor module, was investigated. For this aim, an ethanol injection-based method was applied for liposome and niosome preparation. The syringe-pump device was used for laboratory scale batches production (30 ml for liposomes, 20 ml for niosomes) then a pilot scale (750 ml for liposomes, 1000 ml for niosomes) were obtained using the SPG membrane contactor. Resulted nanovesicles were characterized in terms of mean vesicles size, polydispersity index (PdI) and zeta potential. The drug encapsulation efficiency (E.E.%) was evaluated using two drug-models: caffeine and spironolactone, a hydrophilic and a lipophilic molecule, respectively. As results, nanovectors mean size using the syringe-pump device was comprised between 82 nm and 95 nm for liposomes and between 83 nm and 127 nm for niosomes. The optimal E.E. of caffeine within niosomes, was found around 9.7% whereas the spironolactone E.E. reached 95.6% which may be attributed to its lipophilic properties. For liposomes these values were about 9.7% and 86.4%, respectively. It can be clearly seen that the spironolactone E.E. was slightly higher within niosomes than liposomes. Optimized formulations, which offered smaller size and higher E.E., were selected for pilot scale production using the SPG membrane. It has been found that vesicles characteristics (size and E.E.%) were reproducible using the membrane contactor module. Thus, the current study demonstrated the usefulness of the membrane contactor as a device for scaling-up both liposome and niosome preparations with small mean sizes.

  17. In vitro determination of the efficacy of scorpion venoms as anti-cancer agents against colorectal cancer cells: a nano-liposomal delivery approach.

    Science.gov (United States)

    Al-Asmari, Abdulrahman K; Ullah, Zabih; Al Balowi, Ali; Islam, Mozaffarul

    2017-01-01

    The use of liposomes in biological and medicinal sciences is a relatively new approach. The liposomal strategy greatly depends on the technological advancement in the formation of vesicles of various sizes and properties. In the current study, we encapsulated the venoms obtained from medically important scorpions such as Androctonus bicolor (AB), Androctonus crassicauda (AC), and Leiurus quinquestriatus (LQ). To begin with, our first and foremost aim was to prepare biocompatible and biodegradable nanovesicles. Additionally, we intended to enhance the anti-cancer potential of these encapsulated venoms. The liposomal venoms were prepared by rehydration and dehydration methods. Morphology, particle size, and size distribution of the liposomes were examined by scanning electron microscope (SEM), transmission electron microscope (TEM), and Zetasizer. We found that the prepared liposomes had a smooth surface and a spherical/ovoid shape and existed mainly as single unilamellar vesicles (SUVs). Furthermore, the liposomal formulation of all three venoms exhibited excellent stability and good encapsulation efficiency (EE). Additionally, the anti-cancer potential of the encapsulated venoms was also evaluated on a colorectal cancer cell line (HCT-8). The venom-loaded liposomes showed elevated anti-cancer properties such as low rate of cell survival, higher reactive oxygen species (ROS) generation, and enhancement in the number of apoptotic cells. In addition to this, cell cycle analysis revealed G0/G1 enrichment upon venom treatment. The effect of treatment was more pronounced when venom-liposome was used as compared to free venom on the HCT-8 cell line. Furthermore, we did not observe any interference of liposomal lipids used in these preparations on the progression of cancer cells. Considering these findings, we can conclude that the encapsulated scorpion venoms exhibit better efficacy and act more vigorously as an anti-cancer agent on the colorectal cancer cell line when

  18. Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants.

    Science.gov (United States)

    Panfoli, Isabella; Ravera, Silvia; Podestà, Marina; Cossu, Claudia; Santucci, Laura; Bartolucci, Martina; Bruschi, Maurizio; Calzia, Daniela; Sabatini, Federica; Bruschettini, Matteo; Ramenghi, Luca Antonio; Romantsik, Olga; Marimpietri, Danilo; Pistoia, Vito; Ghiggeri, Gianmarco; Frassoni, Francesco; Candiano, Giovanni

    2016-04-01

    Exosomes are secreted nanovesicles that are able to transfer RNA and proteins to target cells. The emerging role of mesenchymal stem cell (MSC) exosomes as promoters of aerobic ATP synthesis restoration in damaged cells, prompted us to assess whether they contain an extramitochondrial aerobic respiration capacity. Exosomes were isolated from culture medium of human MSCs from umbilical cord of ≥37-wk-old newborns or between 28- to 30-wk-old newborns (i.e.,term or preterm infants). Characterization of samples was conducted by cytofluorometry. Oxidative phosphorylation capacity was assessed by Western blot analysis, oximetry, and luminometric and fluorometric analyses. MSC exosomes express functional respiratory complexes I, IV, and V, consuming oxygen. ATP synthesis was only detectable in exosomes from term newborns, suggestive of a specific mechanism that is not completed at an early gestational age. Activities are outward facing and comparable to those detected in mitochondria isolated from term MSCs. MSC exosomes display an unsuspected aerobic respiratory ability independent of whole mitochondria. This may be relevant for their ability to rescue cell bioenergetics. The differential oxidative metabolism of pretermvs.term exosomes sheds new light on the preterm newborn's clinical vulnerability. A reduced ability to repair damaged tissue and an increased capability to cope with anoxic environment for preterm infants can be envisaged.-Panfoli, I., Ravera, S., Podestà, M., Cossu, C., Santucci, L., Bartolucci, M., Bruschi, M., Calzia, D., Sabatini, F., Bruschettini, M., Ramenghi, L. A., Romantsik, O., Marimpietri, D., Pistoia, V., Ghiggeri, G., Frassoni, F., Candiano, G. Exosomes from human mesenchymal stem cells conduct aerobic metabolism in term and preterm newborn infants.

  19. On-chip immunoelectrophoresis of extracellular vesicles released from human breast cancer cells.

    Directory of Open Access Journals (Sweden)

    Takanori Akagi

    Full Text Available Extracellular vesicles (EVs including exosomes and microvesicles have attracted considerable attention in the fields of cell biology and medicine. For a better understanding of EVs and further exploration of their applications, the development of analytical methods for biological nanovesicles has been required. In particular, considering the heterogeneity of EVs, methods capable of measuring individual vesicles are desired. Here, we report that on-chip immunoelectrophoresis can provide a useful method for the differential protein expression profiling of individual EVs. Electrophoresis experiments were performed on EVs collected from the culture supernatant of MDA-MB-231 human breast cancer cells using a measurement platform comprising a microcapillary electrophoresis chip and a laser dark-field microimaging system. The zeta potential distribution of EVs that reacted with an anti-human CD63 (exosome and microvesicle marker antibody showed a marked positive shift as compared with that for the normal immunoglobulin G (IgG isotype control. Thus, on-chip immunoelectrophoresis could sensitively detect the over-expression of CD63 glycoproteins on EVs. Moreover, to explore the applicability of on-chip immunoelectrophoresis to cancer diagnosis, EVs collected from the blood of a mouse tumor model were analyzed by this method. By comparing the zeta potential distributions of EVs after their immunochemical reaction with normal IgG, and the anti-human CD63 and anti-human CD44 (cancer stem cell marker antibodies, EVs of tumor origin circulating in blood were differentially detected in the real sample. The result indicates that the present method is potentially applicable to liquid biopsy, a promising approach to the low-invasive diagnosis of cancer.

  20. Integrative Nanomedicine: Treating Cancer With Nanoscale Natural Products

    Science.gov (United States)

    Sarter, Barbara; Koithan, Mary; Banerji, Prasanta; Banerji, Pratip; Jain, Shamini; Ives, John

    2014-01-01

    Finding safer and more effective treatments for specific cancers remains a significant challenge for integrative clinicians and researchers worldwide. One emerging strategy is the use of nanostructured forms of drugs, vaccines, traditional animal venoms, herbs, and nutraceutical agents in cancer treatment. The recent discovery of nanoparticles in traditional homeopathic medicines adds another point of convergence between modern nanomedicine and alternative interventional strategies. A way in which homeopathic remedies could initiate anticancer effects includes cell-to-cell signaling actions of both exogenous and endogenous (exosome) nanoparticles. The result can be a cascade of modulatory biological events with antiproliferative and pro-apoptotic effects. The Banerji Protocols reflect a multigenerational clinical system developed by homeopathic physicians in India who have treated thousands of patients with cancer. A number of homeopathic remedy sources from the Banerji Protocols (eg, Calcarea phosphorica; Carcinosin—tumor-derived breast cancer tissue prepared homeopathically) overlap those already under study in nonhomeopathic nanoparticle and nanovesicle tumor exosome cancer vaccine research. Past research on antineoplastic effects of nano forms of botanical extracts such as Phytolacca, Gelsemium, Hydrastis, Thuja, and Ruta as well as on homeopathic remedy potencies made from the same types of source materials suggests other important overlaps. The replicated finding of silica, silicon, and nano-silica release from agitation of liquids in glassware adds a proven nonspecific activator and amplifier of immunological effects. Taken together, the nanoparticulate research data and the Banerji Protocols for homeopathic remedies in cancer suggest a way forward for generating advances in cancer treatment with natural product–derived nanomedicines. PMID:24753994

  1. Extra-cellular release and blood diffusion of BART viral micro-RNAs produced by EBV-infected nasopharyngeal carcinoma cells

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    Baconnais Sonia

    2010-10-01

    Full Text Available Abstract Background Nasopharyngeal carcinoma (NPC is a human epithelial malignancy consistently associated with the Epstein-Barr virus. The viral genome is contained in the nuclei of all malignant cells with abundant transcription of a family of viral microRNAs called BART miRNAs. MicroRNAs are well known intra-cellular regulatory elements of gene expression. In addition, they are often exported in the extra-cellular space and sometimes transferred in recipient cells distinct from the producer cells. Extra-cellular transport of the microRNAs is facilitated by various processes including association with protective proteins and packaging in secreted nanovesicles called exosomes. Presence of microRNAS produced by malignant cells has been reported in the blood and saliva of tumor-bearing patients, especially patients diagnosed with glioblastoma or ovarian carcinoma. In this context, it was decided to investigate extra-cellular release of BART miRNAs by NPC cells and their possible detection in the blood of NPC patients. To address this question, we investigated by quantitative RT-PCR the status of 5 microRNAs from the BART family in exosomes released by NPC cells in vitro as well as in plasma samples from NPC xenografted nude mice and NPC patients. Results We report that the BART miRNAs are released in the extra-cellular space by NPC cells being associated, at least to a large extent, with secreted exosomes. They are detected with a good selectivity in plasma samples from NPC xenografted nude mice as well as NPC patients. Conclusions Viral BART miRNAs are secreted by NPC cells in vitro and in vivo. They have enough stability to diffuse from the tumor site to the peripheral blood. This study provides a basis to explore their potential as a source of novel tumor biomarkers and their possible role in communications between malignant and non-malignant cells.

  2. Smart blood cell and microvesicle-based Trojan horse drug delivery: Merging expertise in blood transfusion and biomedical engineering in the field of nanomedicine.

    Science.gov (United States)

    Wu, Yu-Wen; Goubran, Hadi; Seghatchian, Jerard; Burnouf, Thierry

    2016-04-01

    Therapeutic and diagnostic applications of nanomedicine are playing increasingly important roles in human health. Various types of synthetic nanoparticles, including liposomes, micelles, and other nanotherapeutic platforms and conjugates, are being engineered to encapsulate or carry drugs for treating diseases such as cancer, cardiovascular disorders, neurodegeneration, and inflammations. Nanocarriers are designed to increase the half-life of drugs, decrease their toxicity and, ideally, target pathological sites. Developing smart carriers with the capacity to deliver drugs specifically to the microenvironment of diseased cells with minimum systemic toxicity is the goal. Blood cells, and potentially also the liposome-like micro- and nano-vesicles they generate, may be regarded as ideally suited to perform such specific targeting with minimum immunogenic risks. Blood cell membranes are "decorated" with complex physiological receptors capable of targeting and communicating with other cells and tissues and delivering their content to the surrounding pathological microenvironment. Blood cells, such as erythrocytes, have been developed as permeable carriers to release drugs to diseased tissues or act as biofactory allowing enzymatic degradation of a pathological substrate. Interestingly, attempts are also being made to improve the targeting capacity of synthetic nanoparticles by "decorating" their surface with blood cell membrane receptor-like biochemical structures. Research is needed to further explore the benefits that blood cell-derived microvesicles, as a Trojan horse delivery systems, can bring to the arsenal of therapeutic micro- and nanotechnologies. This short review focuses on the therapeutic roles that red blood cells and platelets can play as smart drug-delivery systems, and highlights the benefits that blood transfusion expertise can bring to this exciting and novel biomedical engineering field.

  3. Interactions between exosomes from breast cancer cells and primary mammary epithelial cells leads to generation of reactive oxygen species which induce DNA damage response, stabilization of p53 and autophagy in epithelial cells.

    Directory of Open Access Journals (Sweden)

    Sujoy Dutta

    Full Text Available Exosomes are nanovesicles originating from multivesicular bodies and are released by all cell types. They contain proteins, lipids, microRNAs, mRNAs and DNA fragments, which act as mediators of intercellular communications by inducing phenotypic changes in recipient cells. Tumor-derived exosomes have been shown to play critical roles in different stages of tumor development and metastasis of almost all types of cancer. One of the ways by which exosomes affect tumorigenesis is to manipulate the tumor microenvironments to create tumor permissive "niches". Whether breast cancer cell secreted exosomes manipulate epithelial cells of the mammary duct to facilitate tumor development is not known. To address whether and how breast cancer cell secreted exosomes manipulate ductal epithelial cells we studied the interactions between exosomes isolated from conditioned media of 3 different breast cancer cell lines (MDA-MB-231, T47DA18 and MCF7, representing three different types of breast carcinomas, and normal human primary mammary epithelial cells (HMECs. Our studies show that exosomes released by breast cancer cell lines are taken up by HMECs, resulting in the induction of reactive oxygen species (ROS and autophagy. Inhibition of ROS by N-acetyl-L-cysteine (NAC led to abrogation of autophagy. HMEC-exosome interactions also induced the phosphorylation of ATM, H2AX and Chk1 indicating the induction of DNA damage repair (DDR responses. Under these conditions, phosphorylation of p53 at serine 15 was also observed. Both DDR responses and phosphorylation of p53 induced by HMEC-exosome interactions were also inhibited by NAC. Furthermore, exosome induced autophagic HMECs were found to release breast cancer cell growth promoting factors. Taken together, our results suggest novel mechanisms by which breast cancer cell secreted exosomes manipulate HMECs to create a tumor permissive microenvironment.

  4. FedExosomes: Engineering Therapeutic Biological Nanoparticles that Truly Deliver

    Directory of Open Access Journals (Sweden)

    Michelle E. Marcus

    2013-04-01

    Full Text Available Many aspects of intercellular communication are mediated through “sending” and “receiving” packets of information via the secretion and subsequent receptor-mediated detection of biomolecular species including cytokines, chemokines, and even metabolites. Recent evidence has now established a new modality of intercellular communication through which biomolecular species are exchanged between cells via extracellular lipid vesicles. A particularly important class of extracellular vesicles is exosomes, which is a term generally applied to biological nanovesicles ~30–200 nm in diameter. Exosomes form through invagination of endosomes to encapsulate cytoplasmic contents, and upon fusion of these multivesicular endosomes to the cell surface, exosomes are released to the extracellular space and transport mRNA, microRNA (miRNA and proteins between cells. Importantly, exosome-mediated delivery of such cargo molecules results in functional modulation of the recipient cell, and such modulation is sufficiently potent to modulate disease processes in vivo. It is possible that such functional delivery of biomolecules indicates that exosomes utilize native mechanisms (e.g., for internalization and trafficking that may be harnessed by using exosomes to deliver exogenous RNA for therapeutic applications. A complementary perspective is that understanding the mechanisms of exosome-mediated transport may provide opportunities for “reverse engineering” such mechanisms to improve the performance of synthetic delivery vehicles. In this review, we summarize recent progress in harnessing exosomes for therapeutic RNA delivery, discuss the potential for engineering exosomes to overcome delivery challenges and establish robust technology platforms, and describe both potential challenges and advantages of utilizing exosomes as RNA delivery vehicles.

  5. Pathologic function and therapeutic potential of exosomes in cardiovascular disease.

    Science.gov (United States)

    Ailawadi, Shaina; Wang, Xiaohong; Gu, Haitao; Fan, Guo-Chang

    2015-01-01

    The heart is a very complex conglomeration of organized interactions between various different cell types that all aid in facilitating myocardial function through contractility, sufficient perfusion, and cell-to-cell reception. In order to make sure that all features of the heart work effectively, it is imperative to have a well-controlled communication system among the different types of cells. One of the most important ways that the heart regulates itself is by the use of extracellular vesicles, more specifically, exosomes. Exosomes are types of nano-vesicles, naturally released from living cells. They are believed to play a critical role in intercellular communication through the means of certain mechanisms including direct cell-to-cell contact, long-range signals as well as electrical and extracellular chemical molecules. Exosomes contain many unique features like surface proteins/receptors, lipids, mRNAs, microRNAs, transcription factors and other proteins. Recent studies indicate that the exosomal contents are highly regulated by various stress and disease conditions, in turn reflective of the parent cell status. At present, exosomes are well appreciated to be involved in the process of tumor and infection disease. However, the research on cardiac exosomes is just emerging. In this review, we summarize recent findings on the pathologic effects of exosomes on cardiac remodeling under stress and disease conditions, including cardiac hypertrophy, peripartum cardiomyopathy, diabetic cardiomyopathy and sepsis-induced cardiovascular dysfunction. In addition, the cardio-protective effects of stress-preconditioned exosomes and stem cell-derived exosomes are also summarized. Finally, we discuss how to epigenetically reprogram exosome contents in host cells which makes them beneficial for the heart.

  6. Modulation of tissue tropism and biological activity of exosomes and other extracellular vesicles: New nanotools for cancer treatment.

    Science.gov (United States)

    Kooijmans, Sander A A; Schiffelers, Raymond M; Zarovni, Natasa; Vago, Riccardo

    2016-09-01

    Exosomes are naturally secreted nanovesicles that have recently aroused a great interest in the scientific and clinical community for their roles in intercellular communication in almost all physiological and pathological processes. These 30-100nm sized vesicles are released from the cells into the extracellular space and ultimately into biofluids in a tightly regulated way. Their molecular composition reflects their cells of origin, may confer specific cell or tissue tropism and underlines their biological activity. Exosomes and other extracellular vesicles (EVs) carry specific sets of proteins, nucleic acids (DNA, mRNA and regulatory RNAs), lipids and metabolites that represent an appealing source of novel noninvasive markers through biofluid biopsies. Exosome-shuttled molecules maintain their biological activity and are capable of modulating and reprogramming recipient cells. This multi-faceted nature of exosomes hold great promise for improving cancer treatment featuring them as novel diagnostic sensors as well as therapeutic effectors and drug delivery vectors. Natural biological activity including the therapeutic payload and targeting behavior of EVs can be tuned via genetic and chemical engineering. In this review we describe the properties that EVs share with conventional synthetic nanoparticles, including size, liposome-like membrane bilayer with customizable surface, and multifunctional capacity. We also highlight unique characteristics of EVs, which possibly allow them to circumvent some limitations of synthetic nanoparticle systems and facilitate clinical translation. The latter are in particular correlated with their innate stability, ability to cross biological barriers, efficiently deliver bioactive cargos or evade immune recognition. Furthermore, we discuss the potential roles for EVs in diagnostics and theranostics, and highlight the challenges that still need to be overcome before EVs can be applied to routine clinical practice.

  7. Improvement of fluvastatin bioavailability by loading on nanostructured lipid carriers

    Directory of Open Access Journals (Sweden)

    El-Helw AR

    2015-09-01

    Full Text Available Abdel-Rahim M El-Helw, Usama A FahmyDepartment of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi ArabiaAbstract: The aim of this study is to prepare fluvastatin nanostructured lipid carriers (FLV-NLCs in order to find an innovative way to alleviate FLV-associated disadvantages. The limitations include poor solubility and extensive first-pass metabolism, resulting in low (30% bioavailability and short elimination half-life (1–3 hours. FLV-NLCs were prepared by hot emulsification–ultrasonication method. Ten runs were created by three-level factorial design (32 to optimize FLV-NLCs formulation process. In this study, two factors, four responses, and three-level factorial design were endorsed. The studied variables were lipid:oil ratio (X1 and sonication time (X2. However, the responses parameter determined the particle size (Y1, nm, entrapment efficiency percent (EE%, Y2, particles zeta potential (Y3, and 80% of the drug release after 24 hours (X4. Furthermore, stability and in vivo pharmacokinetics were studied in rats. The optimized consisted formula had an average particle size of 165 nm with 75.32% entrapment efficiency and 85.32% of drug released after 24 hours, demonstrating a sustaining drug release over 24 hours. An in vivo pharmacokinetic study revealed enhanced bioavailability by >2.64-fold, and the mean residence time was longer than that of FLV. We concluded that NLCs could be promising carriers for sustained/prolonged FLV release with enhanced oral bioavailability.Keywords: factorial design, nano-vesicles, hyperlipidemia, statins

  8. CLL Exosomes Modulate the Transcriptome and Behaviour of Recipient Stromal Cells and Are Selectively Enriched in miR-202-3p.

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    Mosavar Farahani

    Full Text Available Bi-directional communication with the microenvironment is essential for homing and survival of cancer cells with implications for disease biology and behaviour. In chronic lymphocytic leukemia (CLL, the role of the microenvironment on malignant cell behaviour is well described. However, how CLL cells engage and recruit nurturing cells is poorly characterised. Here we demonstrate that CLL cells secrete exosomes that are nanovesicles originating from the fusion of multivesicular bodies with the plasma membrane, to shuttle proteins, lipids, microRNAs (miR and mRNAs to recipient cells. We characterise and confirm the size (50-100 nm and identity of the CLL-derived exosomes by Electron microscopy (EM, Atomic force microscopy (AFM, flow cytometry and western blotting using both exosome- and CLL-specific markers. Incubation of CLL-exosomes, derived either from cell culture supernatants or from patient plasma, with human stromal cells shows that they are readily taken up into endosomes, and induce expression of genes such as c-fos and ATM as well as enhance proliferation of recipient HS-5 cells. Furthermore, we show that CLL exosomes encapsulate abundant small RNAs and are enriched in certain miRs and specifically hsa-miR-202-3p. We suggest that such specific packaging of miR-202-3p into exosomes results in enhanced expression of 'suppressor of fused' (Sufu, a Hedgehog (Hh signalling intermediate, in the parental CLL cells. Thus, our data show that CLL cells secrete exosomes that alter the transcriptome and behaviour of recipient cells. Such communication with microenvironment is likely to have an important role in CLL disease biology.

  9. Interactions between exosomes from breast cancer cells and primary mammary epithelial cells leads to generation of reactive oxygen species which induce DNA damage response, stabilization of p53 and autophagy in epithelial cells.

    Science.gov (United States)

    Dutta, Sujoy; Warshall, Case; Bandyopadhyay, Chirosree; Dutta, Dipanjan; Chandran, Bala

    2014-01-01

    Exosomes are nanovesicles originating from multivesicular bodies and are released by all cell types. They contain proteins, lipids, microRNAs, mRNAs and DNA fragments, which act as mediators of intercellular communications by inducing phenotypic changes in recipient cells. Tumor-derived exosomes have been shown to play critical roles in different stages of tumor development and metastasis of almost all types of cancer. One of the ways by which exosomes affect tumorigenesis is to manipulate the tumor microenvironments to create tumor permissive "niches". Whether breast cancer cell secreted exosomes manipulate epithelial cells of the mammary duct to facilitate tumor development is not known. To address whether and how breast cancer cell secreted exosomes manipulate ductal epithelial cells we studied the interactions between exosomes isolated from conditioned media of 3 different breast cancer cell lines (MDA-MB-231, T47DA18 and MCF7), representing three different types of breast carcinomas, and normal human primary mammary epithelial cells (HMECs). Our studies show that exosomes released by breast cancer cell lines are taken up by HMECs, resulting in the induction of reactive oxygen species (ROS) and autophagy. Inhibition of ROS by N-acetyl-L-cysteine (NAC) led to abrogation of autophagy. HMEC-exosome interactions also induced the phosphorylation of ATM, H2AX and Chk1 indicating the induction of DNA damage repair (DDR) responses. Under these conditions, phosphorylation of p53 at serine 15 was also observed. Both DDR responses and phosphorylation of p53 induced by HMEC-exosome interactions were also inhibited by NAC. Furthermore, exosome induced autophagic HMECs were found to release breast cancer cell growth promoting factors. Taken together, our results suggest novel mechanisms by which breast cancer cell secreted exosomes manipulate HMECs to create a tumor permissive microenvironment.

  10. Proteomic analysis of exosomes from nasopharyngeal carcinoma cell identifies intercellular transfer of angiogenic proteins.

    Science.gov (United States)

    Chan, Yuk-Kit; Zhang, Huoming; Liu, Pei; Tsao, Sai-Wah; Lung, Maria Li; Mak, Nai-Ki; Ngok-Shun Wong, Ricky; Ying-Kit Yue, Patrick

    2015-10-15

    Exosomes, a group of secreted extracellular nanovesicles containing genetic materials and signaling molecules, play a critical role in intercellular communication. During tumorigenesis, exosomes have been demonstrated to promote tumor angiogenesis and metastasis while their biological functions in nasopharyngeal carcinoma (NPC) are poorly understood. In this study, we focused on the role of NPC-derived exosomes on angiogenesis. Exosomes derived from the NPC C666-1 cells and immortalized nasopharyngeal epithelial cells (NP69 and NP460) were isolated using ultracentrifugation. The molecular profile and biophysical characteristics of exosomes were verified by Western blotting, sucrose density gradient and electron microscopy. We showed that the C666-1 exosomes (10 and 20 μg/ml) could significantly increase the tubulogenesis, migration and invasion of human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner. Subsequently, an iTRAQ-based quantitative proteomics was used to identify the differentially expressed proteins in C666-1 exosomes. Among the 640 identified proteins, 51 and 89 proteins were considered as up- and down-regulated (≥ 1.5-fold variations) in C666-1 exosomes compared to the normal counterparts, respectively. As expected, pro-angiogenic proteins including intercellular adhesion molecule-1 (ICAM-1) and CD44 variant isoform 5 (CD44v5) are among the up-regulated proteins, whereas angio-suppressive protein, thrombospondin-1 (TSP-1) was down-regulated in C666-1 exosomes. Further confocal microscopic study and Western blotting clearly demonstrated that the alteration of ICAM-1 and TSP-1 expressions in recipient HUVECs are due to internalization of exosomes. Taken together, these data strongly indicated the critical roles of identified angiogenic proteins in the involvement of exosomes-induced angiogenesis, which could potentially be developed as therapeutic targets in future.

  11. Hypoxic enhancement of exosome release by breast cancer cells

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    King Hamish W

    2012-09-01

    Full Text Available Abstract Background Exosomes are nanovesicles secreted by tumour cells which have roles in paracrine signalling during tumour progression, including tumour-stromal interactions, activation of proliferative pathways and bestowing immunosuppression. Hypoxia is an important feature of solid tumours which promotes tumour progression, angiogenesis and metastasis, potentially through exosome-mediated signalling. Methods Breast cancer cell lines were cultured under either moderate (1% O2 or severe (0.1% O2 hypoxia. Exosomes were isolated from conditioned media and quantitated by nanoparticle tracking analysis (NTA and immunoblotting for the exosomal protein CD63 in order to assess the impact of hypoxia on exosome release. Hypoxic exosome fractions were assayed for miR-210 by real-time reverse transcription polymerase chain reaction and normalised to exogenous and endogenous control genes. Statistical significance was determined using the Student T test with a P value of  Results Exposure of three different breast cancer cell lines to moderate (1% O2 and severe (0.1% O2 hypoxia resulted in significant increases in the number of exosomes present in the conditioned media as determined by NTA and CD63 immunoblotting. Activation of hypoxic signalling by dimethyloxalylglycine, a hypoxia-inducible factor (HIF hydroxylase inhibitor, resulted in significant increase in exosome release. Transfection of cells with HIF-1α siRNA prior to hypoxic exposure prevented the enhancement of exosome release by hypoxia. The hypoxically regulated miR-210 was identified to be present at elevated levels in hypoxic exosome fractions. Conclusions These data provide evidence that hypoxia promotes the release of exosomes by breast cancer cells, and that this hypoxic response may be mediated by HIF-1α. Given an emerging role for tumour cell-derived exosomes in tumour progression, this has significant implications for understanding the hypoxic tumour phenotype, whereby hypoxic

  12. Biosolar energy generation and harvesting from biomolecule-copolymer hybrid systems

    Science.gov (United States)

    Chu, Bong-Chieh Benjamin

    Alternative energy sources have become an increasingly important topic as energy needs outpace supply. Furthermore, as the world moves into the digital age of portable electronics, highly efficient and lightweight energy sources will need to be developed. Current technology, such as lithium ion batteries, provide enough power to run portable electronics for hours or days, but can still allow for improvement in their power density (W/kg). Utilizing energy-transducing membrane proteins, which are by nature highly efficient, it is possible to engineer biological-based energy sources with energy densities far greater than any solid-state systems. Furthermore, solar powered membrane proteins have the added benefit of a virtually unlimited supply of energy. This work has developed protein-polymer hybrid films and nanoscale vesicles for a variety of applications from fuel-cell technology to biological-based photovoltaics. Bacteriorhodopsin (BR), a light-activated proton pump, and Cytochrome C Oxidase (COX), a protein involved in the electron transport chain in mitochondria, were reconstituted into biomimetic triblock copolymer membranes. Block copolymer membranes mimic the amphiphilic nature of a natural lipid bilayer but exhibit greater mechanical stability due to UV-polymerizable endgroups. In BR/COX functionalized nanovesicles, proton gradients generated by the light-activated proton pumping of BR are used to drive COX in reverse to generate electrons, providing a hybrid biologically-active polymer to convert solar energy to chemical energy, and finally to electrical energy. This work has found protein activity in planar membranes through the photoelectric current generation by BR and the proton pumping activity of BR-functionalized polymer membranes deposited onto proton exchange membranes, as well as the coupled functionality of BR and COX through current generation in cyclic voltammetry and direct current measurements. Current switching between light and dark

  13. Connective tissue growth factor (CCN2) and microRNA-21 are components of a positive feedback loop in pancreatic stellate cells (PSC) during chronic pancreatitis and are exported in PSC-derived exosomes.

    Science.gov (United States)

    Charrier, Alyssa; Chen, Ruju; Chen, Li; Kemper, Sherri; Hattori, Takako; Takigawa, Masaharu; Brigstock, David R

    2014-06-01

    Pancreatitis is an inflammatory condition of the pancreas which, in its chronic form, involves tissue destruction, exocrine and endocrine insufficiency, increased risk of pancreatic cancer, and an extensive fibrotic pathology which is due to unrelenting collagen deposition by pancreatic stellate cells (PSC). In response to noxious agents such as alcohol-excessive consumption of which is a major cause of pancreatitis in the West-normally quiescent PSC undergo a phenotypic and functional transition to activated myofibroblasts which produce and deposit collagen at high levels. This process is regulated by connective tissue growth factor (CCN2), expression of which is highly up-regulated in activated PSC. We show that CCN2 production by activated PSC is associated with enhanced expression of microRNA-21 (miR-21) which was detected at high levels in activated PSC in a murine model of alcoholic chronic pancreatitis. A positive feedback loop between CCN2 and miR-21 was identified that resulted in enhancement of their respective expression as well as that of collagen α1(I). Both miR-21 and CCN2 mRNA were present in PSC-derived exosomes, which were characterized as 50-150 nm CD9-positive nano-vesicles. Exosomes from CCN2-GFP- or miR-21-GFP-transfected PSC were taken up by other PSC cultures, as shown by direct fluorescence or qRT-PCR for GFP. Collectively these studies establish miR-21 and CCN2 as participants in a positive feedback loop during PSC activation and as components of the molecular payload in PSC-derived exosomes that can be delivered to other PSC. Thus interactions between cellular or exosomal miR-21 and CCN2 represent novel aspects of fibrogenic regulation in PSC. Summary Chronic injury in the pancreas is associated with fibrotic pathology which is driven in large part by CCN2-dependent collagen production in pancreatic stellate cells. This study shows that CCN2 up-regulation in PSC is associated with increased expression of miR-21 which, in turn, is able to

  14. Infrared spectroscopic study of thermotropic phase behavior of newly developed synthetic biopolymers.

    Science.gov (United States)

    Bista, Rajan K; Bruch, Reinhard F; Covington, Aaron M

    2011-10-15

    The thermotropic phase behavior of a suite of newly developed self-forming synthetic biopolymers has been investigated by variable-temperature Fourier transform infrared (FT-IR) absorption spectroscopy. The temperature-induced infrared spectra of these artificial biopolymers (lipids) composed of 1,2-dimyristoyl-rac-glycerol-3-dodecaethylene glycol (GDM-12), 1,2-dioleoyl-rac-glycerol-3-dodecaethylene glycol (GDO-12) and 1,2-distearoyl-rac-glycerol-3-triicosaethylene glycol (GDS-23) in the spectral range of 4000-500 cm(-1) have been acquired by using a thin layered FT-IR spectrometer in conjunction with a custom built temperature-controlled demountable liquid cell having a pathlength of ∼15 μm. The lipids under consideration have long hydrophobic acyl chains and contain various units of hydrophilic polyethylene glycol (PEG) headgroups. In contrast to conventional phospholipids, this new kind of lipids forms liposomes or nanovesicles spontaneously upon hydration, without requiring external activation energy. We have found that the thermal stability of the PEGylated lipids differs greatly depending upon the acyl chain-lengths as well as the nature of the associated bonds and the number of PEG headgroup units. In particular, GDM-12 (saturated 14 hydrocarbon chains with 12 units of PEG headgroup) exhibits one sharp order-disorder phase transition over a temperature range increasing from 3°C to 5°C. Similarly, GDS-23 (saturated 18 hydrocarbon chains with 23 units of PEG headgroup) displays comparatively broad order-disorder phase transition profiles between temperature 17°C and 22°C. In contrast, GDO-12 (monounsaturated 18 hydrocarbon chains with 12 units of PEG headgroup) does not reveal any order-disorder transition phenomena demonstrating a highly disordered behavior for the entire temperature range. To confirm these observations, differential scanning calorimetry (DSC) was applied to the samples and revealed good agreement with the infrared spectroscopy results

  15. Hierarchical assemblies of soft matters from polymers and liquid crystals on structured surfaces

    Science.gov (United States)

    Honglawan, Apiradee

    Hierarchical, multifunctional materials hold important keys to numerous advanced technologies, including electronics, optics, and medicine. This thesis encompasses generation of hierarchical structures with novel morphologies and functions through self-assembly directed by lithographically fabricated templates. Here, two soft materials, amphiphilic random copolymers of photopolymerized acryloyl chloride (ranPAC) and smectic-A liquid crystal (SmA-LC) molecule, 4'(5,5,6,6,7,7,8,8,9,9,10,10,11,11,12,12,12-heptadecaflu-orododecyloxy)-biphenyl-4-carboxylic acid ethyl ester, are synthesized as model systems to investigate the governing principles at the topographic surface/interface. The ranPAC can self-organize into nanomicelles with high regularity and stability, typically not possible in random copolymer systems. The morphology can be controlled by the photopolymerization conditions and solvent; the crosslinked shell makes the micelles robust against drying and storage. Using SU-8 micropillar arrays with spatially controlled surface chemistry as templates, we construct hierarchical microporous structures with tunable pore size and symmetry (e.g. square array), and uncover a new evaporative assembly method. By functionalizing the ranPAC nanovesicles with cationic poly(ethyleneimines), we encapsulate the anticancer drug, doxorubicin hydrochloride, and mRNA at a high payload, which are delivered to HEK 293T cells in vitro at a low cytotoxicity level. SmA-LC are characterized by arrangement of molecules into thin layers with the long molecular axis parallel to the layer normal, forming a close-packed hexagonal array of topological defects known as focal conic domains (FCDs) in a thin film. Using a series of SU-8 micropillar arrays with different size, shape, height, and symmetry as topological templates, we investigate the epitaxial and hierarchical assemblies of FCDs; whether the system favors confinement or "pillar edge-pinning" depends on balance of the elastic energy

  16. Trans-nasal zolmitriptan novasomes: in-vitro preparation, optimization and in-vivo evaluation of brain targeting efficiency.

    Science.gov (United States)

    Abd-Elal, Radwa M A; Shamma, Rehab N; Rashed, Hassan M; Bendas, Ehab R

    2016-11-01

    Migraine attack is a troublesome physiological condition associated with throbbing, intense headache, in one half of the head. Zolmitriptan is a potent second-generation triptan, prescribed for patients with migraine attacks, with or without an aura, and cluster headaches. The absolute bioavailability of zolmitriptan is about 40% for oral administration; due to hepatic first metabolism. Nasal administration would circumvent the pre-systemic metabolism thus increasing the bioavailability of zolmitriptan. In addition, due to the presence of microvilli and high vasculature, the absorption is expected to be faster compared to oral route. However, the bioavailability of nasal administered drugs is particularly restricted by poor membrane penetration. Thus, the aim of this work is to explore the potential of novel nanovesicular fatty acid enriched structures (novasomes) for effective and enhanced nasal delivery of zolmitriptan and investigate their nose to brain targeting potential. Novasomes were prepared using nonionic surfactant, cholesterol in addition to a free fatty acid. A 2(3) full factorial design was adopted to study the influence of the type of surfactant, type of free fatty acid and ratio between the free fatty acid and the surfactant on novasomes properties. The particle size, entrapment efficiency, polydispersity index, zeta potential and % zolmitriptan released after 2 h were selected as dependent variables. Novasomes were further optimized using Design Expert® software (version 7; Stat-Ease Inc., Minneapolis, MN), and an optimized formulation composed of Span® 80:Cholesterol:stearic acid (in the ratio 1:1:1) was selected. This formulation showed zolmitriptan entrapment of 92.94%, particle size of 149.9 nm, zeta potential of -55.57 mV, and released 48.43% zolmitriptan after 2 h. The optimized formulation was further examined using transmission electron microscope, which revealed non-aggregating multi-lamellar nanovesicles with narrow size

  17. 外质体(Exosomes)与肾脏疾病%Exosomes and kidney diseases

    Institute of Scientific and Technical Information of China (English)

    柏云

    2012-01-01

    Exosomes are nanovesicles originating from multivesicular bodies ( MVBs) and secreted into the extracellular space or body fluids when a multivesicular body {endocytic origin) fuses with the plasma membrane. Exosomes contain multiple proteins, mRNAs, microRNAs, and signaling molecules that may reflect the physiological state of their cells of origin and consequently provide potential biomarkers. At present,the studies on exosomes are mostly focused on their roles in immunology and oncology and exosorne-based immunotherapy has become a new means in cancer treatment and immune tolerance. In recent years, urinary exosomes (UE) and their roles in kidney diseases have been receiving great attention. Exosomes are secreted to the urine from all types of renal epithelial cell, including glomerular podocytes, renal tubular cells, and the cells lining the urinary drainage system. Thus, urinary exosomes have potential as a source of valuable biomarkers for early detection of kidney diseases. The present review aims to summarize their biological characteristics,and their potential uses in the diagnosis and treatment of kidney disease.%外质体( Exosomes)足起源于多泡体的微小囊泡,由细胞内吞途径中的多泡体外膜和细胞膜融合后释放到胞外环境或体液中.Exosomes含有多种蛋白、mRNAs、microRNAs、信号分子等,能够反映来源细胞的生物学状态,因而可能成为潜在的生物学标志物.目前,exosomes的研究大多集中在免疫学和肿瘤学,并已经成为一种免疫治疗的新手段,应用于肿瘤治疗和免疫耐受等方面.近年人们才关注exosomes与肾脏疾病的关系,研究表明几乎所有肾脏上皮细胞包括肾小球足细胞、肾小管上皮细胞、尿道上皮细胞均可分泌exosomes,因此尿液来源的exosomes可能成为寻找肾脏疾病早期诊断的标志物.本文着重从exosomes的生物学特性及其在肾脏疾病诊断和治疗的研究进行综述.

  18. Thin nanostructured crystalline TiO{sub 2} films and their applications in solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Cheng Yajun

    2007-06-15

    morphology of TiO{sub 2} films can also be controlled. The local and long range structures of the titania films were investigated by the combination of imaging techniques (AFM, SEM) and x-ray scattering techniques (X-ray reflectivity and grazing incidence small-angle X-ray scattering). Based on the knowledge of the morphology control, the crystalline TiO{sub 2} nanostructured films with different morphologies were introduced into solid state dye-sensitized solar cells. It has been found that all of the morphologies help to improve the performance of the solar cells. Especially, clustered nanoparticles, worm-like structures, foam-like structures, large collapsed nanovesicles show more pronounced performance improvement than other morphologies such as nanowires, flakes, and nanogranulars. (orig.)

  19. Preparation and Characterization of Self-assembly Vesicle of Icariin Nanofiber%淫羊藿苷纳米纤维膜自组装囊泡的制备及表征

    Institute of Scientific and Technical Information of China (English)

    江永南; 莫红缨

    2012-01-01

    OBJECTIVE To improve the compatibility and absorption properties of icariin in vivo by preparing electrospun icariin nanofiber and forming self-assembly nano-vesicles. METHODS Solubility test in solvent was performed to screen suitable solvent system. Icariin nanofibers was prepared by electrospinning. SEM and TEM were used to observe fiber membrane surface and self-assembly vehicle morphology respectively. XRD, DSC and IR were applied to characterize the drug-loaded nanofilber. RESULTS Methanol and dimethylacetamide were selected as the solvent mixture for providing good solubility and formation of drug-loaded fiber. The electron scan microscope showed that diameter of fiber was (400-600 nm) and the fiber surface was smooth without drug absorption. The polymers were well compatible with drug. Hydrogen bonds played a key interaction between drug and polymers. Nano-vehicle was self-assembled by dispersing the drug-loaded nanofiler into water. CONCLUSION Preparation of icariin nanofiber membrane by electrospinning was simple and icariin was highly dispersed in nanofiberse in amorphous form. Nanofibers can be self-assembled in water to form nano-vehicles.%目的 采用静电纺丝制备淫羊藿苷纳米纤维膜,并通过自组装技术形成纳采囊泡,改善淫羊藿苷在体内的相容性及吸收性能.方法 通过溶解度实验筛选合适溶剂,以静电纺丝技术制备淫羊藿苷纳米纤维膜,采用扫描电镜对纤维膜表面形态进行观察,采用X射线晶体衍射(XRD)和差示扫描量热分析(DSC)检测纤维膜中药物的存在状态,通过红外光谱分析药物与纤维材料之间的相互作用.并通过透射电镜观察纳米纤维膜自组装纳采囊泡的性能.结果 甲醇与二甲基乙酰胺混合溶剂的溶解性及纤维成型性较好;载药纤维直径分布均匀(400~600 nm)、表面光滑无药物颗粒,药物与聚合物之间通过氢键作用,具有良好的相容性,水中溶解试验发现纳米纤维膜能

  20. CBIOS Science Sessions - 2016 - Part I and III National Symposium on Nanoscience and Biomedical Nanotechnology - Proceedings

    Directory of Open Access Journals (Sweden)

    L. Monteiro Rodrigues, et al.

    2016-05-01

    nanophysics | Nanoquímica e nanofísica Chairman | Moderador - José Paulo Farinha Speakers | Prelectores João Pedro Conde João Rodrigues Beatriz Nobre 1st Session | Sessão 1 Nanomedicine | Nanomedicina Chairman / Moderador Catarina Pinto Reis C.01 - Across the blood-brain barrier: Nanoparticles Passagem através da barreira hematoencefálica: Nanopartículas Speaker / Prelector Jorg Kreuter C.02 - Nanovesicles for dermal and transdermal delivery Nanovesículas para administração dérmica e transdérmica Speaker / Prelector Sandra Simões C.03 - Ligand-functionalized nanoparticles for treatment of melanoma in situ Nanopartículas funcionalizadas com ligandos para o tratamento in situ do melanoma Poster selected for oral communication / Poster selecionado para comunicação oral Catarina Silva Chairman / Moderador Luís P. Fonseca C.04 - Targeted and local delivery of nucleic acids Administração vectorizada e local de ácidos nucleicos Speaker / Prelector Elias Fattal C.05 - Gold precision: Gold nanoparticles for precise delivery Precisão de ouro: Nanopartículas de ouro para administração precisa Speaker / Prelector Pedro Viana Baptista C.06 - Phytosome as leading strategy for natural compounds delivery Fitossomas como estratégias de administração de compostos naturais Poster selected for oral communication / Poster selecionado para comunicação oral Diogo Matias 2st Session | Sessão 2 Biomedical nanotechnologies | Nanotecnologia Biomédica Chairman / Moderador Hugo Alexandre Ferreira C.07 - Functionalisation of gold nanoparticles for biomedical application Funcionalização de nanopartículas de ouro para aplicação biomédica Speaker / Prelector Sónia Fraga C.08 - Light-activatable biomaterials for biomedical applications Biomateriais fotoactivados para aplicações biomédicas Speaker / Prelector Lino Ferreira C.09 - 7α-acetoxy-6β-hidroxyroyleanone derivatives and its topical application through delivery nanosystems Derivados da 7α -acetoxi-6