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1.
Nat Commun ; 11(1): 4536, 2020 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-32913189

RESUMO

Natural musculoskeletal systems have been widely recognized as an advanced robotic model for designing robust yet flexible microbots. However, the development of artificial musculoskeletal systems at micro-nanoscale currently remains a big challenge, since it requires precise assembly of two or more materials of distinct properties into complex 3D micro/nanostructures. In this study, we report femtosecond laser programmed artificial musculoskeletal systems for prototyping 3D microbots, using relatively stiff SU-8 as the skeleton and pH-responsive protein (bovine serum albumin, BSA) as the smart muscle. To realize the programmable integration of the two materials into a 3D configuration, a successive on-chip two-photon polymerization (TPP) strategy that enables structuring two photosensitive materials sequentially within a predesigned configuration was proposed. As a proof-of-concept, we demonstrate a pH-responsive spider microbot and a 3D smart micro-gripper that enables controllable grabbing and releasing. Our strategy provides a universal protocol for directly printing 3D microbots composed of multiple materials.


Assuntos
Biomimética/métodos , Compostos de Epóxi/efeitos da radiação , Fenômenos Fisiológicos Musculoesqueléticos , Polímeros/efeitos da radiação , Robótica/métodos , Soroalbumina Bovina/efeitos da radiação , Biomimética/instrumentação , Compostos de Epóxi/química , Hidrogéis/química , Hidrogéis/efeitos da radiação , Concentração de Íons de Hidrogênio , Lasers , Polimerização/efeitos da radiação , Polímeros/química , Impressão Tridimensional , Robótica/instrumentação , Soroalbumina Bovina/química
2.
Int J Nanomedicine ; 15: 5459-5471, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32801700

RESUMO

Purpose: Indocyanine green (ICG), a near infrared (NIR) dye clinically approved in medical diagnostics, possesses great heat conversion efficiency, which renders itself as an effective photosensitizer for photothermal therapy (PTT) of cancer. However, there remain bottleneck challenges for use in PTT, which are the poor photo and plasma stability of ICG. To address these problems, in this research, ICG-loaded silver nanoparticles were prepared and evaluated for the applicability as an effective agent for photothermal cancer therapy. Methods and Results: PEGylated bovine serum albumin (BSA)-coated silver core/shell nanoparticles were synthesized with a high loading of ICG ("PEG-BSA-AgNP/ICG"). Physical characterization was carried out using size analyzer, transmission electron microscopy, and Fourier transform infrared spectrophotometry to identify successful preparation and size stability. ICG-loading content and the photothermal conversion efficiency of the particles were confirmed with inductively coupled plasma mass spectrometry and laser instruments. In vitro studies showed that the PEG-BSA-AgNP/ICG could provide great photostability for ICG, and their applicability for PTT was verified from the cellular study results. Furthermore, when the PEG-BSA-AgNP/ICG were tested in vivo, study results exhibited that ICG could stably remain in the blood circulation for a markedly long period (plasma half-life: 112 min), and about 1.7% ID/g tissue could be accumulated in the tumor tissue at 4 h post-injection. Such nanoparticle accumulation in the tumor enabled tumor surface temperature to be risen to 50°C (required for photo-ablation) by laser irradiation and led to successful inhibition of tumor growth in the B16F10 s.c. syngeneic nude mice model, with minimal systemic toxicity. Conclusion: Our findings demonstrated that PEG-BSA-AgNPs could serve as effective carriers for delivering ICG to the tumor tissue with great stability and safety.


Assuntos
Antineoplásicos/farmacologia , Nanopartículas Metálicas/química , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/química , Animais , Antineoplásicos/efeitos adversos , Antineoplásicos/farmacocinética , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos/métodos , Estabilidade de Medicamentos , Difusão Dinâmica da Luz , Meia-Vida , Humanos , Verde de Indocianina/química , Verde de Indocianina/farmacocinética , Melanoma Experimental/tratamento farmacológico , Camundongos Endogâmicos ICR , Camundongos Nus , Microscopia Eletrônica de Transmissão , Polietilenoglicóis/química , Soroalbumina Bovina/química , Prata/química , Espectroscopia de Infravermelho com Transformada de Fourier
3.
J Chromatogr A ; 1628: 461443, 2020 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-32822982

RESUMO

Sodium dodecyl sulfate (SDS) in proteomics samples needs to be removed and estimated prior to mass spectrometry (MS)-based analysis and to avoid MS ion-source contamination. Here, we describe an organic solvent free method to remove SDS using a simple apparatus that mainly consists of an agarose gel inside a 1 mL plastic micropipette tip and a voltage power supply with electrodes. A small volume of sample (e.g., 50 µL) is loaded on top of the gel and then voltage (cathode at the sample side) is applied with an acidic solution at the other end of the micropipette tip. Within 25 min, SDS was removed (e.g., ≥99% SDS in 3.5 mM SDS) and the peptides were retained in the sample solution. The strategy was compared to the commercially available and expensive Pierce spin column for the removal of SDS and recovery of peptides from a digested bovine serum albumin sample.


Assuntos
Técnicas de Química Analítica/métodos , Técnicas Eletroquímicas , Proteômica/métodos , Dodecilsulfato de Sódio/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Espectrometria de Massas , Peptídeos/química , Soroalbumina Bovina/química , Dodecilsulfato de Sódio/química
4.
J Chromatogr A ; 1628: 461444, 2020 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-32822983

RESUMO

The effect of bead and ligand structure on protein adsorption was investigated for multimodal anion exchangers combining a quaternary ammonium ion group with hydrophobic moieties: Nuvia aPrime 1 and aPrime 2, based on a 54 µm diameter polymeric bead, and Capto Adhere ImpRes and Capto Adhere, based on agarose beads 51 and 78 µm diameter, respectively. Bovine serum albumin (BSA) monomer, BSA dimer, and thyroglobulin (Tg) were used as model proteins. Based on TEM imaging and iSEC, the Nuvia resins have a microgranular structure and large pores (110 nm radius), while the Capto resins have a fibrous structure and smaller pores (32-36 nm radius). Comparable binding capacities (80-110 mg/mL), decreasing as salt is added, are observed for all three proteins on the Nuvia resins. Higher capacities (110-130 mg/mL), also decreasing as salt is added, are observed for BSA monomer and dimer on the Capto resins. However, the Tg binding capacity is very low in this case and increases as salt is added. Confocal laser scanning microscopy show that the kinetics are controlled by pore diffusion for all four resins, but with diffusivities that decrease as the protein size increases especially for the Capto resins. For Tg at low salt, binding is restricted to a thin shell close to the bead surface for both Capto resins. The ratio of effective and free diffusivity is about 0.30, 0.18, and 0.08 for BSA monomer, BSA dimer, and Tg, respectively, on the Nuvia resin. These values decrease to about 0.11, 0.04, and 0.01, respectively, for the Capto resins as a result of diffusional hindrance. Dynamic binding capacities are consistent with the equilibrium and rate behaviors.


Assuntos
Resinas de Troca de Ânions/química , Cromatografia por Troca Iônica , Proteínas/metabolismo , Adsorção , Ânions/química , Difusão , Interações Hidrofóbicas e Hidrofílicas , Cinética , Ligantes , Polímeros/química , Sefarose/química , Soroalbumina Bovina/química
5.
Int J Nanomedicine ; 15: 5203-5215, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32801686

RESUMO

Introduction: Metformin is an ideal candidate to treat the liver tumor with insulin resistance because of its good performance in the treatment of type 2 diabetes and the advantage in cancer therapy. We aim to develop a delivery system with higher efficiency than free drug. Methods: Metformin-bovine serum albumin (met-BSA) nanoparticles (NPs) were prepared using the anti-solvent precipitation method with a stabilizer of BSA for particle growth. The therapeutic effect of the drug was tested by the insulin-resistant HepG2 cells and C57BL/6J mice at a glucose starvation condition. The interaction mechanism of the drug and the protein during the formation of the NPs was tested using a series of spectroscopy. Results: Metformin and BSA formed nonporous and spherical particles of about 200 nm with proper lognormal distribution and thermostability. The cellular uptake, as well as the anti-liver cancer activities of met-BSA, was enhanced dramatically compared with the free drug. The thermodynamic studies suggested that the weak binding of metformin to BSA was governed by hydrogen bonds and van der Waals forces. Moreover, the results of synchronous, circular dichroism (CD) and three-dimensional fluorescence demonstrated that the BSA skeleton and chromophore microenvironments were changed in the presence of metformin. Conclusion: Therefore, met-BSA has been proved as a simple yet effective therapeutic agent for cancer with insulin resistance, promising for future clinic translations in cancer treatment.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Resistência à Insulina , Metformina/farmacologia , Nanopartículas/administração & dosagem , Soroalbumina Bovina/farmacologia , Animais , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/patologia , Dicroísmo Circular , Diabetes Mellitus Tipo 2 , Células Hep G2 , Humanos , Ligação de Hidrogênio , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/patologia , Masculino , Metformina/administração & dosagem , Metformina/química , Camundongos Endogâmicos C57BL , Nanopartículas/química , Soroalbumina Bovina/química , Termodinâmica , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Int J Nanomedicine ; 15: 4171-4189, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32606671

RESUMO

Background: Angiogenic and osteogenic activities are two major problems with biomedical titanium (Ti) and other orthopedic implants used to repair large bone defects. Purpose: The aim of this study is to prepare hydroxyapatite (HA) coatings on the surface of Ti by using electrochemical deposition (ED), and to evaluate the effects of nanotopography and silicon (Si) doping on the angiogenic and osteogenic activities of the coating in vitro. Materials and Methods: HA coating and Si-doped HA (HS) coatings with varying nanotopographies were fabricated using two ED modes, ie, the pulsive current (PC) and cyclic voltammetry (CV) methods. The coatings were characterized through scanning electron microscope (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectrometer (XPS), and atomic force microscopy (AFM), and their in vitro bioactivity and protein adsorption were assessed. Using MC3T3-E1 pre-osteoblasts and HUVECs as cell models, the osteogenic and angiogenic capabilities of the coatings were evaluated through in vitro cellular experiments. Results: By controlling Si content in ~0.8 wt.%, the coatings resulting from the PC mode (HA-PC and HS-PC) and CV mode (HA-CV and HS-CV) had nanosheet and nanorod topographies, respectively. At lower crystallinity, higher ionic dissolution, smaller contact angle, higher surface roughness, and more negative zeta potential, the HS and PC samples exhibited quicker apatite deposition and higher BSA adsorption capacity. The in vitro cell study showed that Si doping was more favorable for enhancing the viability of the MC3T3-E1 cells, but nanosheet coating increased the area for cell spreading. Of the four coatings, HS-PC with Si doping and nanosheet topography exhibited the best effect in terms of up-regulating the expressions of the osteogenic genes (ALP, Col-I, OSX, OPN and OCN) in the MC3T3-E1 cells. Moreover, all leach liquors of the surface-coated Ti disks promoted the growth of the HUVECs, and the HS samples played a more significant role in promoting cell migration and tube formation than the HA samples. Of the four leach liquors, only the two HS samples up-regulated NO content and expressions of the angiogenesis-related genes (VEGF, bFGF and eNOS) in the HUVECs, and the HS-PC yielded a better effect. Conclusion: The results show that Si doping while regulating the topography of the coating can help enhance the bone regeneration and vascularization of HA-coated Ti implants.


Assuntos
Materiais Revestidos Biocompatíveis/farmacologia , Durapatita/farmacologia , Nanopartículas/química , Osteogênese , Próteses e Implantes , Silício/química , Titânio/farmacologia , Adsorção , Animais , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Camundongos , Nanopartículas/ultraestrutura , Nanotubos/química , Neovascularização Fisiológica/efeitos dos fármacos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Soroalbumina Bovina/química , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Difração de Raios X
7.
Food Chem ; 333: 127432, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32659661

RESUMO

The sensory qualities and shelf life of tea beverage strongly affected by tea cream that forms by the interaction of polyphenols and protein. The study aimed to investigate the effects of the interactions between tea polyphenols (TPs) and bovine serum albumin (BSA) on tea cream formation at different concentrations. The tea cream formation increased with TPs and BSA concentration increased. The optimal concentration (TPs: 800 mg/L, BSA: 40 mg/L), for high clarities and contents of phytochemicals, was selected by the technique for order preference by similarity to ideal solution (C = 0.7572). The interaction mechanism of TPs-BSA was investigated by fluorescence spectroscopy, UV-visible absorption spectroscopy, synchronous fluorescence spectroscopy, and molecular docking. TPs interacted with BSA via static quenching process, affecting tryptophan and tyrosine residue microenvironment of BSA. Ester catechins had more binding affinity than non-ester catechins. Hydrogen bonds were the main interaction forces of TPs-BSA.


Assuntos
Polifenóis/química , Polifenóis/metabolismo , Soroalbumina Bovina/química , Chá/química , Animais , Sítios de Ligação , Catequina/química , Catequina/metabolismo , Precipitação Química , Ligação de Hidrogênio , Simulação de Acoplamento Molecular , Tamanho da Partícula , Soroalbumina Bovina/metabolismo , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Chá/metabolismo , Termodinâmica
8.
J Chromatogr A ; 1625: 461277, 2020 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-32709329

RESUMO

The influence of impregnation the chromatographic plate adsorbent layer, silica, with hen's egg white albumin (OVA) or bovine serum albumin (BSA) on the retention of some popular medicines (paracetamol, aminophenazone, theophylline, caffeine, acetanilide, ciprofloxacin, tramadol, acetylsalicylic acid, acebutolol) is investigated. The effect of composition and buffer pH of the mobile phase on solute separation selectivity is also studied. The chromatographic systems with and without above mentioned albumins and their influence on investigated drug retention are compared. In general, it has been turned out that retention of tested medicines in systems with the sorbent impregnated with albumin significantly increase relative to those with non-impregnated.


Assuntos
Cromatografia em Camada Delgada/métodos , Clara de Ovo/química , Preparações Farmacêuticas/análise , Soroalbumina Bovina/química , Sílica Gel/química , Adsorção , Animais , Bovinos , Galinhas , Feminino , Concentração de Íons de Hidrogênio , Metanol/química , Compostos Orgânicos/química , Solventes/química , Espectroscopia de Infravermelho com Transformada de Fourier , Tolueno/química
9.
J Chromatogr A ; 1625: 461284, 2020 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-32709334

RESUMO

In this work, a class of novel and eco-friendly open-tubular (OT) chiral column was presented for the first time by one step preparation with zeolite imidazolate framework-8 (ZIF-8) and bovine serum album (BSA) based on electrostatic adsorption and adsorption affinity. This stationary phase materials combined the features of large surface areas and adsorption affinity of ZIF-8, and also the multiple chiral binding sites of BSA, which contributes to the π-interaction and hydrophobic interaction with the analytes. The separation performance of BSA@ZIF-8-OT chiral columns was evaluated with a miniaturized capillary electrochromatography and amperometric detection (mini-CEC-AD) system; in particular, nine groups of model molecules, including homologues, structural isomers, and chiral compounds, were baseline separated under the certain optimum conditions. The RSDs of run-to-run, day-to-day, column-to-column, and batch-to-batch reproducibility were less than 13.8 %. Furthermore, the prepared OT columns were successfully applied to fast analysis of ephedrine isomers in Chinese herb ephedra, and the LODs achieved 1.5-2.0 ng mL-1 (S/N=3) by an electrophoretic stacking technique of moving chemical reaction boundary. This mini-CEC-AD system with BSA@ZIF-8-OT chiral columns provides a promising potential in pharmaceutical analysis due to its fast, sensitive, enantioselective, and low-cost characteristics.


Assuntos
Eletrocromatografia Capilar/métodos , Imidazóis/química , Soroalbumina Bovina/química , Zeolitas/química , Adsorção , Animais , Bovinos , Concentração de Íons de Hidrogênio , Limite de Detecção , Reprodutibilidade dos Testes , Estereoisomerismo
10.
PLoS One ; 15(7): e0235263, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32639981

RESUMO

Dependent peptide searching is a method for discovering covalently-modified peptides-and therefore proteins-in mass-spectrometry-based proteomics experiments. Being more permissive than standard search methods, it has the potential to discover novel modifications (e.g., post-translational modifications occurring in vivo, or modifications introduced in vitro). However, few studies have explored dependent peptide search results in an untargeted way. In the present study, we sought to evaluate dependent peptide searching as a means of characterising proteins that have been modified in vitro. We generated a model data set by analysing N-ethylmaleimide-treated bovine serum albumin, and performed dependent peptide searches using the popular MaxQuant software. To facilitate interpretation of the search results (hundreds of dependent peptides), we developed a series of visualisation tools (R scripts). We used the tools to assess the diversity of putative modifications in the albumin, and to pinpoint hypothesised modifications. We went on to explore the tools' generality via analyses of public data from studies of rat and human proteomes. Of 19 expected sites of modification (one in rat cofilin-1 and 18 across six different human plasma proteins), eight were found and correctly localised. Apparently, some sites went undetected because chemical enrichment had depleted necessary analytes (potential 'base' peptides). Our results demonstrate (i) the ability of the tools to provide accurate and informative visualisations, and (ii) the usefulness of dependent peptide searching for characterising in vitro protein modifications. Our model data are available via PRIDE/ProteomeXchange (accession number PXD013040).


Assuntos
Peptídeos/análise , Proteínas/química , Proteômica/métodos , Animais , Proteínas Sanguíneas/química , Bovinos , Bases de Dados de Proteínas , Etilmaleimida/análogos & derivados , Humanos , Processamento de Proteína Pós-Traducional , Ratos , Soroalbumina Bovina/química
11.
Food Chem ; 328: 127122, 2020 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-32480260

RESUMO

Glycation between target proteins and saccharides is time-consuming or requires high temperatures. Here, a promising reaction medium, natural deep eutectic solvents (NADES), for glucose glycation with bovine serum albumin (BSA) was applied to improve the grafting of glucose-glycated BSA by shifting reaction equilibrium. Two types of glucose-glycated BSA products were prepared using NADES and water systems. SDS-PAGE and MALDI-TOF-MS revealed that BSA and glucose were covalently bonded. Compared with in water system, glycated BSA products in NADES system had more -OH groups, more disordered secondary structures, lower intrinsic fluorescence intensity, and higher ultraviolet-visible absorption. Lower surface hydrophobicity (1100 versus 1356), higher emulsifying activity index (66.17 versus 46.49 m2/g), higher emulsion stability index (79.62 versus 63.61%), and lesser free sulfhydryl (8.07 versus 8.98 µmol/g) groups were obtained with NADES system than with water system. The results suggest that NADES is a suitable alternative reaction medium for promoting the glycation of BSA.


Assuntos
Glucose/química , Soroalbumina Bovina/química , Animais , Glicosilação , Interações Hidrofóbicas e Hidrofílicas , Solventes/química
12.
Food Chem ; 330: 127170, 2020 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-32531633

RESUMO

With the increasing concerns on food safety, foodborne toxicants, particularly advanced glycation end products (AGEs) have drawn large attention ascribing to their central role in the pathogenic process of diabetic complications. In this study, the antiglycative ability and action mechanism of 10 dried flowers were measured in fructose/glucose-BSA model with apple flower as the most effective one, probably through trapping reactive carbonyl species. Furthermore, apple flower was added in cookies to evaluate its antiglycative effect and data suggested that the fortification of cookies with apple flower powders significantly inhibited the formation of methylglyoxal and fluorescent AGEs. Moreover, sensory evaluation data showed that 2.5% (w/w) apple flower-fortified cookies had the highest acceptance. The active component in apple flower was further identified as phlorizin by chromatographic analysis. Our results highlighted the health benefits of consuming apple flower fortified foods which might possibly be developed into functional foods especially for diabetic patients.


Assuntos
Frutose/química , Glucose/química , Produtos Finais de Glicação Avançada/química , Malus/química , Animais , Flores/química , Aldeído Pirúvico/química , Soroalbumina Bovina/química
13.
Artigo em Inglês | MEDLINE | ID: mdl-32515655

RESUMO

To understand the effect of 17α-ethinylestradiol (EE2) on the conformation changes of bovine serum albumin (BSA), the binding mechanisms of EE2 with BSA were investigated by fluorescence spectroscopy, time-resolved fluorescence spectroscopy, synchronous fluorescence spectroscopy, three-dimensional fluorescence spectroscopy, UV-visible spectroscopy, circular dichroism (CD) spectroscopy and molecular docking. The quenching constants, binding constants, the number of binding sites, thermodynamic parameters, binding distance and the secondary structure changes of BSA were determined. The results of fluorescence quenching experiment suggested that the fluorescence quenching of BSA by EE was due to the formation of complex through static quenching, which was also confirmed by time-resolved fluorescence measurements. The thermodynamic parameters indicated that the binding of EE2 to BSA was driven mainly by van der Waals forces and hydrogen bonding. The conformation alterations of BSA upon EE2 binding were studied by UV-vis spectroscopy and CD spectroscopy. The results of site marker competitive experiments and molecular docking showed that the binding sites of EE2 were mainly located within site I in the subdomain IIA of BSA.


Assuntos
Dicroísmo Circular/métodos , Poluentes Ambientais/química , Etinilestradiol/química , Soroalbumina Bovina/química , Espectrometria de Fluorescência/métodos , Espectrofotometria Ultravioleta/métodos , Animais , Sítios de Ligação , Bovinos , Simulação de Acoplamento Molecular , Ligação Proteica , Conformação Proteica , Termodinâmica
14.
Environ Toxicol ; 35(11): 1170-1178, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32519799

RESUMO

Formaldehyde (FA) can be produced in the environment and by cell metabolism and has been classified as a carcinogen in animals and humans. Metformin is the most commonly used drug for the treatment of type 2 diabetes. Metformin also has potential benefit in cancer prevention and treatment. The aim of this study was to determine whether metformin can directly react with FA and attenuate its toxicity in vitro. Metformin was incubated at pH 7.4 and 37°C in the presence of FA, and the reaction mixture was analyzed by UV spectrophotometry, high-performance liquid chromatography (HPLC), and mass spectrometry. Fluorescence spectrophotometry, immunofluorescence, and western blot were used to measure FA-induced bovine serum albumin (BSA) crosslinking and DNA damage in HepG2 cells treated with or without metformin. According to the HPLC and mass spectrometry data, we speculate that the reaction of metformin with FA (1:1) initially results in the formation of a conjugated intermediate followed by the subsequent generation of a stable six-membered ring structure. Correspondingly, metformin attenuated FA-induced fluorescence in BSA as well as the aggregation of γH2AX in HepG2 cells. These results suggest that metformin can protect protein and DNA damage induced by FA at least partly through a direct reaction process.


Assuntos
Formaldeído/toxicidade , Metformina/metabolismo , Animais , Dano ao DNA/efeitos dos fármacos , Humanos , Soroalbumina Bovina/química
15.
Int J Nanomedicine ; 15: 3539-3550, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32547012

RESUMO

Background: Methotrexate (MTX) is an antiproliferative drug widely used to treat inflammatory diseases and autoimmune diseases. The application of percutaneous administration is hindered due to its poor transdermal penetration. To reduce side effects and enhanced percutaneous delivery of MTX, novel methotrexate (MTX)-loaded micelles prepared with a amphiphilic cationic material, N,N-dimethyl-(N',N'-di-stearoyl-1-ethyl)1,3-diaminopropane (DMSAP), was designed. Materials and Methods: DMSAP was synthesized via three steps using simple chemical agents. H nuclear magnetic resonance and mass spectroscopy were used to confirm the successful synthesis of DMSAP. A safe and non-toxic phosphatidylcholine, soybean phosphatidylcholine (SPC), was added to DMSAP at different ratios to form P/D-micelles. Then, MTX-entrapped micelles (M/P/D-micelles) were prepared by electrostatic adsorption. The physicochemical properties and blood stability of micelles were examined thoroughly. In addition, the transdermal potential of the micelles was evaluated by permeation experiments. Results: In aqueous environments, DMSAP conjugates could self-assemble spontaneously into micelles with a low critical micelle concentration (CMC) of 0.056 mg/mL. Stable, spherical MTX-entrapped micelles (M/P/D-micelles) with a size of 100-120 nm and high zeta potential of +36.26 mV were prepared. In vitro permeation studies showed that M/P/D-micelles exhibited superior skin permeability and deposition of MTX in the epidermis and dermis compared with that of free MTX. Conclusion: These special novel cationic M/P/D-micelles can enhance the permeability of MTX and are expected to be a promising percutaneous delivery system for therapy skin diseases.


Assuntos
Metotrexato/administração & dosagem , Micelas , Administração Cutânea , Animais , Cátions , Bovinos , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Humanos , Metotrexato/química , Camundongos , Concentração Osmolar , Tamanho da Partícula , Fosfatidilcolinas/química , Espectroscopia de Prótons por Ressonância Magnética , Soroalbumina Bovina/química , Pele/efeitos dos fármacos , Eletricidade Estática
16.
Int J Nanomedicine ; 15: 3433-3445, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32523342

RESUMO

Background: Reconstituted lipoproteins (rLips) based on endogenous lipid nanostructures has been increasingly regarded as an excellent and promising antitumor drug delivery. However, some problems relating to the main component, apolipoprotein, for instance, rare source, unaffordable price, and low specificity of relevant receptor expression, become chief obstacles to its broad development and application. Purpose: The primary aim of this study is to develop biomimetic rLips by utilizing folic acid (FA)-modified bovine serum albumin (BSA) as a replacement for apolipoprotein and demonstrate its tumor targeting and antitumor efficacy. Methods: The amino groups of BSA were covalently conjugated with FA through the amide reaction. PTX-loaded nanostructured lipid carrier (termed as P-NLC) consisting of phospholipid, cholesteryl ester, triglyceride and cholesterol was prepared by the emulsification-evaporation method and utilized as the lipid core. FA-modified BSA (FA-BSA) was characterized for the protein substitute degree and attached with NLC by incubation-insert method to form the lipoprotein-mimic nanocomplex (termed as PFB-rLips). The morphology of nanoparticles was observed under transmission electron microscopy (TEM), and the particle size and zeta potential were determined using dynamic light scattering. In vitro release behavior of PTX from PFB-rLips was investigated with the dialysis method. Hemolysis tests were conducted to evaluate the biosecurity of PFB-rLips. Cell uptake and cytotoxicity assays were performed on human hepatocytes (LO2) and human hepatoma cells (HepG2). Tumor targeting was assessed using in vivo imaging system in H22 tumor-bearing mice model. Antitumor efficacy in vivo was investigated and compared between Taxol® (paclitaxel) formulation and PTX-incorporated nanoparticles in the same tumor model. Results: A fixed molar ratio 50:1 of FA to BSA was chosen as the optimal input ratio based on the balance between appropriate degree of protein substitution and amphiphilicity of FA-BSA. The morphology of FB-rLips exhibited as a homogeneous spherical structure featured by lipid cores surrounded with a cloudy protein shell observed under TEM. The particle size, zeta potential and encapsulation efficiency were 174.6±3.2 nm, -17.26±0.9 mV and 82.2±2.4%, respectively. In vitro release behavior of PTX from PFB-rLips was slow and sustained. The uptake of FB-rLips was much higher in HepG2 cells than in LO2 cells. Furthermore, the uptake of FB-rLips was significantly higher than that of rLips without FA involved (termed as B-rLips) and NLC in HepG2 cells. Hemolysis and cytotoxicity assays showed good biocompatibility of FB-rLips. The internalization mechanism of FB-rLips mainly depended on clathrin-mediated and caveolin-mediated endocytosis coupling with energy consumption, and FA receptors expressed on tumor cells played a critical role in cellular uptake process. CCK-8 studies demonstrated that PFB-rLips exhibited significantly better tumor killing ability than Taxol® (paclitaxel) formulation in vitro. Moreover, FB-rLips produced more excellent tumor-targeting properties than NLC through in vivo imaging assays. On the basis of this, PTX-loaded FB-rLips also performed more remarkable anticancer activity than other therapy groups in H22 tumor-bearing mice. Conclusion: FB-rLips would serve as a potential nanocarrier for improving tumor-targeting and therapeutic efficacy while reducing the side effects on normal tissues and organs.


Assuntos
Portadores de Fármacos/química , Ácido Fólico/uso terapêutico , Lipoproteínas/química , Nanopartículas/química , Neoplasias/tratamento farmacológico , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Liberação Controlada de Fármacos , Endocitose/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Ácido Fólico/síntese química , Ácido Fólico/química , Hemólise/efeitos dos fármacos , Humanos , Camundongos , Nanopartículas/ultraestrutura , Neoplasias/patologia , Paclitaxel/química , Paclitaxel/farmacologia , Paclitaxel/uso terapêutico , Tamanho da Partícula , Coelhos , Soroalbumina Bovina/química , Eletricidade Estática
17.
Chemosphere ; 259: 127440, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32590178

RESUMO

A new mixed matrix membrane (MMM) was prepared by incorporating biological mesoporous carbon microspheres (mCMSs) from corn starch polysaccharide-supported hydrophilic polydopamine (PDA), as a mesoporous and large-surface area filler, selective modifier, and pore-forming agent, into polyvinylidene fluoride (PVDF) matrix in presence of polyethylene glycol (PEG) as a hydrophilic agent. The structural parameters of the prepared membranes were characterized via FE-SEM, BET/BJH, XRD, FT-IR, and AFM analyses, sorption experiments, water permeability assessments, porosimetry tests, flux recovery ratio (FRR) evaluations, and contact angle measurements, with the so-called central composite design (CCD) been successfully applied for optimization and investigation of the effects of the operational parameters. The results were then applied to treat double-distilled water containing bovine serum albumin (BSA) utilizing a cross-module set-up. Based on the findings, the content of the mCMS-PDA in the PVDF matrix significantly affected the contact angle, pure water flux (PWF), FRR, and BSA removal. In this respect, the PWF of the PVDF-PEG-mCMS-PDA increased from 10.25 to 27.78 L/m2 h with increasing the mCMS-PDA content, with the peak FRR (93.84%) of the PVDF-PEG-mCMS-PDA seen at maximum surface hydrophilicity of the membrane.


Assuntos
Carbono/química , Membranas Artificiais , Soroalbumina Bovina/química , Purificação da Água/métodos , Interações Hidrofóbicas e Hidrofílicas , Indóis , Microesferas , Permeabilidade , Polímeros , Polivinil , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química , Zea mays
18.
AAPS PharmSciTech ; 21(5): 164, 2020 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-32488630

RESUMO

The aim of present research work was to design, fabricate, optimize, and evaluate allopurinol (ALLO)-loaded bovine serum albumin nanoparticles (ABNPs) for kidney targeting of the drug and exploring the potential of fabricated ABNPs for management of hyperuricemia-related nephrolithiasis. ABNP formulation was prepared by employing desolvation technique, and its optimization was conducted by 2-factor-3-level central composite design (CCD) in order to achieve minimum particle size (PSA) and polydispersity index (PDI), maximum entrapment efficiency (EE), and zeta potential (ZP). Further, the optimized formulation (ABNPsopt) was also assessed for in vitro drug release study, TEM, DSC, XRD analysis, FTIR spectroscopy, and in vivo animal study. The in vivo study revealed that after 2 h of ABNPsopt administration, a significant concentration of ALLO was present in kidney (21.26-fold) as compared with serum while in case of standard pure drug group; no drug was seen in mice kidney and serum post 2 h administration, which indicates successful targeting of ALLO by formulating its albumin nanoparticles. Also, uric acid and pH levels were measured in serum and urine samples of mice which showed significant (P < 0.01) efficacy of ABNPsopt formulation in management of hyperuricemia-related nephrolithiasis. Histological studies on kidney samples also confirmed these outcomes. Findings of present study indicate higher kidney uptake of allopurinol from formulated ABNPsopt, which could be due to the specificity of albumin polymer for cubilin and megalin receptors, and it also serves as effective strategy in management of hyperuricemic-related nephrolithiasis.


Assuntos
Alopurinol/administração & dosagem , Supressores da Gota/administração & dosagem , Hiperuricemia/tratamento farmacológico , Nefrolitíase/tratamento farmacológico , Soroalbumina Bovina/química , Alopurinol/uso terapêutico , Animais , Portadores de Fármacos/química , Composição de Medicamentos , Supressores da Gota/uso terapêutico , Humanos , Hiperuricemia/complicações , Rim , Camundongos , Nanopartículas/química , Nefrolitíase/complicações , Tamanho da Partícula
19.
J Chromatogr A ; 1621: 461079, 2020 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-32360061

RESUMO

Capto™ Core 700 is a core-shell chromatographic support with an adsorbing core contained within an inert shell layer designed to purify larger biomolecules and bioparticles in a flow-through mode. The present study aims to characterize the structure and functional properties of this resin using bovine serum albumin (BSA, Mr~65 kDa) and thyroglobulin (Tg, Mr~660 kDa) as model impurity proteins. The functionalized adsorbing core and the inert shell have the same fibrous structure typical of agarose-based beads. The resin average bead size is 90.7 µm with a range of 50-130 µm, the shell thickness is 4.18 µm with a range of 3-6 µm and a standard deviation of 0.55 µm, and the pore radius, obtained by inverse size exclusion chromatography, is 50.4 ± 1.3 nm. Both proteins present highly favorable binding isotherms with maximum binding capacities of 55 and 105 mg/mL of total bead volume for BSA and Tg, respectively. The addition of 500 mM NaCl reduces the binding capacity by less than 50%, showing the ability of the resin to operate at high salt conditions. For both proteins, the effective pore diffusivity in the core is smaller than in the shell due to additional hindrance by bound protein in the core area. Effective pore diffusivities values in the core are 1.6 × 10-7 and 0.16 × 10-7 cm2/s for BSA and Tg, respectively. The DBC10% at 2 min residence time are 24 and 2 mg/mL for BSA and Tg, respectively. This study provides qualitative and quantitative information about Capto™ Core 700 resin. This information could be used to predict and optimize the purification of large biomolecules and bioparticle in route to the establishment of more effective downstream processes.


Assuntos
Cromatografia em Gel/métodos , Tamanho da Partícula , Adsorção , Animais , Bovinos , Resinas Sintéticas/química , Soroalbumina Bovina/química , Temperatura , Tireoglobulina/química
20.
Soft Matter ; 16(21): 5044-5053, 2020 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-32452496

RESUMO

Conventional approaches to mitigate fouling of membrane surfaces impart hydrophilicity to the membrane surface, which increases the water of hydration and fluidity near the surface. By contrast, we demonstrate here that tuning the membrane surface energy close to that of the dispersive component of water surface tension (21.8 mN m-1) can also improve the antifouling properties of the membrane. Specifically, ultrafiltration (UF) membranes were first modified using polydopamine (PDA) followed by grafting of amine-terminated polysiloxane (PSi-NH2). For example, with 2 g L-1 PSi-NH2 coating solution, the obtained coating layer contains 53% by mass fraction PSi-NH2 and exhibits a total surface energy of 21 mN m-1, decreasing the adsorption of bovine serum albumin by 44% compared to the unmodified membrane. When challenged with 1 g L-1 sodium alginate in a constant-flux crossflow system, the PSi-NH2-grafted membrane exhibits a 70% lower fouling rate than the pristine membrane at a water flux of 110 L (m2 h)-1 and good stability when cleaned with NaOH solutions.


Assuntos
Incrustação Biológica/prevenção & controle , Membranas Artificiais , Siloxanas/química , Absorção Fisico-Química , Alginatos/química , Indóis/química , Polímeros/química , Soroalbumina Bovina/química , Ultrafiltração/métodos , Purificação da Água/métodos
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